CN108490194A - Detect the homogeneous immunological detection reagent suit and its preparation method and application of follicular stimulating hormone - Google Patents
Detect the homogeneous immunological detection reagent suit and its preparation method and application of follicular stimulating hormone Download PDFInfo
- Publication number
- CN108490194A CN108490194A CN201810142754.9A CN201810142754A CN108490194A CN 108490194 A CN108490194 A CN 108490194A CN 201810142754 A CN201810142754 A CN 201810142754A CN 108490194 A CN108490194 A CN 108490194A
- Authority
- CN
- China
- Prior art keywords
- stimulating hormone
- chamber
- follicular stimulating
- receptor
- concentration
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/76—Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Endocrinology (AREA)
- Hematology (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Reproductive Health (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to the homogeneous immunological detection reagent of a kind of detection follicular stimulating hormone of biotechnology suits and its preparation method and application.The homogeneous immunological detection reagent suit of detection follicular stimulating hormone prepared by the method for the invention, when being analyzed using light-induced chemiluminescent immunoassay platform, high sensitivity, precision are good, and by optimizing analysis system (antibody dosage and buffer solution etc.), analytical performance meets professional standard, and the associated disease for clinical diagnosis follicular stimulating hormone horizontal abnormality provides reference.
Description
Technical field
The invention belongs to biotechnologies, and in particular to a kind of homogeneous immunological detection reagent set of detection follicular stimulating hormone
Dress and its preparation method and application.
Background technology
Follicular stimulating hormone is also known as follicle-stimulating hormone (FSH) (Follicle-stimulating hormone, FSH), be one kind by brain
The hormone that hypophysis is synthesized and secreted is the glycosylated protein of heterodimer.Follicular stimulating hormone is by two polypeptide groups of α and β
At, and have with glycoprotein hormones such as H-TSH (TSH), lutropin (LH), human chorionic gonadtropins (hCG)
There are identical α subunits, and β subunits are then different with the difference of hormone, wherein β subunits are responsible for mutual with follicle-stimulating hormone receptor
Effect.Follicular stimulating hormone is all critically important one of hormone in men and women's hermaphroditism.In women body, follicular stimulating hormone stimulates still
The growth of immature ovarian follicle, until ripe is Graafian follicle.Ovarian follicle can discharge inhibin to block ovum in growth course
Steep the further synthesis of stimulin.This mechanism ensure that the selectivity of ovulation.At the end in leuteinization stage, follicular stimulating hormone
Level also has to be promoted by a small margin, may be related with the beginning of next ovulatory cycle.In males, follicular stimulating hormone improves plug
Er Tuoli cells synthesize the protein-bonded level of male sex hormone, induce combining closely for Sertoli cell, while secreting inhibition
Element plays the role of vital at Process of Spermatozoa.In clinic, the exception of follicular stimulating hormone, which increases, to be common in：It is primary
Property amenorrhoea, the illnesss such as primary hypogona dism, early stage anterior pituitary hyperfunction, seminoma of testis；And ovarian follicle stimulates
Plain exception is lowly common in：Estrogen or progesterone treatment secondary hypogonadism, Skien syndrome (also known as seat Chinese synthesis
Disease) drugs such as late period primary pituitary hypofunction and intake oral contraceptive sex hormone.
The method of detection follicular stimulating hormone includes enzyme-linked immunosorbent assay, radioactive immunoassay, chemiluminescence at present
Immunoassay etc..Enzymatic activity is affected by many factors in enzyme-linked immunosorbent assay, poor repeatability, and operation every time is both needed to make mark
Directrix curve can increase the testing cost of single part of sample, meanwhile, " hook effect " is easy tod produce, error detection result is caused；Radiation
The major defect of immunoassay is the environmental pollution that radionuclide is brought, and radioactively labelled substance half-life short, kit
Shelf life is short, is not easy to store.Three of the above chemiluminescence immunoassay method has common although marker used is different
Feature belongs to heterogeneous immunoassay (Heterogeneous immunoassay).Heterogeneous immunoassay is believed in detection
It before number, needs by washing solid phase (particle) repeatedly, the labelled antibody for not participating in reaction in liquid phase is distributed in separation removal.
Although solid phase adsorption separation method has many merits such as simple, quick, repeatedly washing will necessarily be immunoassay band
" error " is washed, due to washing process especially EIA enzyme immunoassay, it is difficult to realize standardization, effect is washed between each instrument connection
Fruit difference, influences the precision detected to a certain extent, occurs between criticizing, batch interior difference, so that impact analysis method is accurate
Degree and analysis susceptibility.
Therefore, there is an urgent need for research and develop a kind of "dead" pollution and reproducible, precision, accuracy and sensitivity at present
High follicular stimulating hormone detection method.
Invention content
In order to solve the above technical problems, the present invention, which provides a kind of prepare, detects follicular stimulating hormone (FSH) homogeneous immune detection
The follicular stimulating hormone in homogeneous immunization method detection sample to be tested can be used in reagent set, the reagent set obtained by this method,
Entire detection process not only saves detection time, and not will produce additional waste liquid without separating, washing process, exempts washing error,
With higher precision, accuracy and sensitivity, analytical performance meets professional standard and clinical labororatory requires.
For this purpose, first aspect present invention provides a kind of homogeneous immunological detection reagent suit preparing detection follicular stimulating hormone
Method comprising：
First chamber is prepared, the first chamber includes component a and the first buffer solution, and wherein component a is and the
What one combining unit combined can react the receptor for generating detectable signal with singlet oxygen；First combining unit can be with
First epitope specificity of follicular stimulating hormone combines；
Second chamber is prepared, the second chamber includes component b and the second buffer solution, and wherein component b is and the
The second combining unit that one marker combines；Second combining unit can be with the second epitope specificity knot of follicular stimulating hormone
It closes, and second epitope and the first epitope non-overlapping；
Third composition is prepared, the third composition includes component c and third buffer solution, wherein the component c is
What is combined with the specific junction mixture of the first marker can generate the donor of singlet oxygen in excited state.
First marker of the present invention and the first label combination are not limited to biotin and Streptavidin.
According to the present invention, first combining unit and the second combining unit be selected from polyclonal antibody, monoclonal antibody,
Antibody binding fragment, artificial antibody, modification antibody, be preferably selected from monoclonal antibody and/or polyclonal antibody.
In certain embodiments of the present invention, in the first chamber component a a concentration of 0.2~0.8mg/mL；
Preferably, in the first chamber component a a concentration of 0.4~0.6mg/mL.
In other embodiments of the present invention, a concentration of 2~8 μ g/mL of component b in the second chamber；It is excellent
Selection of land, a concentration of 4~6 μ g/mL of component b in the second chamber.
In certain embodiments of the present invention, in the third composition component c a concentration of 50~150 μ g/mL；It is excellent
Selection of land, a concentration of 80~120 μ g/mL of component c in the third composition.
According to invention, the method further includes preparing follicular stimulating hormone calibration object serial dilutions.
In certain specific embodiments of the invention, a concentration of the 0 of the follicular stimulating hormone calibration object serial dilutions
~200mIU/mL；Preferably, the concentration of the follicular stimulating hormone calibration object serial dilutions be respectively 0mIU/mL, 1mIU/mL,
10mIU/mL, 50mIU/mL, 100mIU/mL and 200mIU/mL.
In certain embodiments of the present invention, the preparation method of the first chamber includes：
Receptor is diluted to 3~8mg/mL using carbonate buffer solution, obtains receptor solution by step S1；
The first combining unit is added in receptor solution, is added after static and is diluted to 9 with carbonate buffer solution by step S2
The BSA solution of~10mg/mL obtains the receptor solution combined with the first combining unit；
Step S3 detaches the receptor combined with the first combining unit in the receptor solution combined with the first combining unit,
And the first buffer solution is added, obtain first chamber.
In other embodiments of the present invention, the preparation method of the second chamber includes：
Second combining unit is placed in bag filter and is dialysed using label buffer solution, added after dialysis by step T1
Enter the first marker solution, and fills into elution buffer, it is static, obtain the second combining unit combined with the first marker；
The second combining unit combined with the first marker is placed in bag filter and is carried out using elution buffer by step T2
The second buffer solution is added in dialysis after dialysis, obtain second chamber solution.
In certain embodiments of the present invention, the NaHCO that the label buffer solution is 0.09~0.1M3Solution；It is described
Elution buffer is the Tris-HCl solution of 0.09~0.1M.
In other embodiments of the present invention, the molar ratio of second combining unit and the first marker is 1:
(20~40).
According to the present invention, first buffer solution and the second buffer solution be that pH value is 7.0~8.5 0.09~
The Tris-HCl solution of 0.10M.
In certain embodiments of the present invention, the receptor includes olefin(e) compound and metallo-chelate, is non-grain
Sub- form, and it is solvable in water-bearing media；And/or the receptor is the macromolecule filled with luminophor and lanthanide series
Particle；
In other embodiments of the present invention, the donor is photoactivation or chemical activation sensitizer, is
Non- particulate forms, and it is solvable in water-bearing media；And/or the donor is the high molecular particle filled with Photoactive compounds.
Second aspect of the present invention provides a kind of detection follicular stimulating hormone prepared by method as described in the first aspect of the invention
Homogeneous immunological detection reagent suit.
Third aspect present invention provides a kind of using reagent set detection ovarian follicle thorn as described in respect of the second aspect of the invention
The homogeneous immunization method of hormone, the method detect for light-induced chemiluminescent.
In certain embodiments of the present invention, the method includes：
Sample to be tested is mixed with first chamber and second chamber, obtains the first mixture by step R1；
First mixture is mixed with third composition, obtains the second mixture by step R2；
Step R3 excites the donor in the second mixture to generate singlet oxygen using energy or reactive compound, described
Receptor can be reacted with the singlet oxygen received generates detectable chemiluminescence signal；
The presence of the chemiluminescence signal obtained in step R4, detecting step R3 and/or intensity wait for test sample to judge to survey
With the presence or absence of the content of follicular stimulating hormone and/or determining follicular stimulating hormone in product.
In other embodiments of the present invention, the method further includes：It is dilute using follicular stimulating hormone calibration object series
Release the step of liquid makes the standard curve of chemiluminescence signal-follicular stimulating hormone concentration；The standard curve is to be measured for determining
The content of follicular stimulating hormone in sample.
In the present invention, after all reagent combinations, mixing can be carried out as needed and/or incubates (incubation).Specifically,
The temperature of the incubation can be 35-45 DEG C, and the time can be 5-30min；Preferably, the temperature of the incubation can be selected from 36
DEG C, 37 DEG C, 38 DEG C, 39 DEG C, 40 DEG C, 41 DEG C or 42 DEG C；The time of incubation can be selected from 10min, 12min, 15min, 16min,
18min, 20min, 25min are stayed overnight.
In certain specific embodiments of the invention, it the described method comprises the following steps：
A. the sample to be tested of 10~30 μ L is added in reacting hole；
B. second chamber described in first chamber and 10~30 μ L described in 10~30 μ L is sequentially added in reacting hole,
A concentration of 0.2~0.8mg/mL of component a in middle first chamber, a concentration of 2~8 μ g/ of the component b in second chamber
mL；
C.35~45 DEG C incubation 10~30 minutes；
D. 100~250 μ L of the third composition are added into reacting hole；Component c's wherein in third composition is dense
Degree is 50~150 μ g/mL；
E.35~45 DEG C incubation 5~20 minutes；
F. it is the laser irradiation reacting hole of 680nm to utilize wavelength, and excited donor generates singlet oxygen, receptor and is touched
Singlet oxygen reaction produces the transmitting light of 612nm；
G. the photon amount for detecting transmitting light in each reacting hole, the concentration of follicular stimulating hormone is calculated according to standard curve.
Here, it should be strongly noted that the above method is the method for non-disease diagnostic purpose.
Fourth aspect present invention provides a kind of reagent set as described in respect of the second aspect of the invention and is preparing for detecting
Application in the kit of follicular stimulating hormone horizontal abnormality associated disease；Preferably, the illness include primary amenorrhea, it is primary
Property hypogona dism, early stage anterior pituitary hyperfunction, seminoma of testis, estrogen or the secondary sexual gland work(of progesterone treatment
It can decline and Skien syndrome late period primary pituitary hypofunction.
Beneficial effects of the present invention are：The homogeneous immune detection examination of detection follicular stimulating hormone prepared by the method for the invention
Agent is set with, and when being analyzed using light-induced chemiluminescent immunoassay platform, high sensitivity, precision are good, and passes through optimization point
Analysis system (antibody dosage and buffer solution etc.), analytical performance meets professional standard, is clinical diagnosis follicular stimulating hormone horizontal abnormality
Associated disease provide reference.
Description of the drawings
To make the present invention be readily appreciated that, illustrate the present invention below in conjunction with the accompanying drawings.
Fig. 1 is the reaction mechanism figure for the homogeneous immunological detection reagent suit for detecting follicular stimulating hormone；Wherein：1 anti-FSH-antibody
In conjunction with luminous microballoon；2 anti-FSH-antibodies combined with biotin；3 FSH to be measured；
The 4 photosensitive microballoons combined with Streptavidin.
Fig. 2 is the dependency diagram of the measurement result and Beckman definite value of embodiment 2.
Specific implementation mode
To make the present invention be readily appreciated that, the present invention is described more detail below.But before describing the present invention in detail, it should be understood that
The present invention is not limited to the specific implementation modes of description.It is also understood that term used herein is embodied only for description
Mode, and be not offered as restrictive.
In the case where providing numberical range, it should be understood that in the upper and lower bound of the range and the prescribed limit
Any other regulation or between two parties each of between numerical value numerical value is encompassed by the present invention between two parties.These small range of upper limits
It can independently be included in smaller range with lower limit, and be also covered by within the present invention, obey any clear in prescribed limit
The limit of exclusion.Defined range include one or two limit in the case of, exclude any of the limit that those include or
The range of the two is also included in the present invention.
Unless otherwise defined, all terms used herein and those skilled in the art's is usual
Understand meaning having the same.Although similar or equivalent any method and material also may be used with method described herein and material
To be used in the implementation or test of the present invention, but preferred method and material will now be described.
I, terms
English corresponding to term " homogeneous " of the present invention is defined as " homogeneous ", and referring to need not be to combining
Antigen antibody complex and remaining free antigen or antibody carry out separation and can be detected.
Term " sample to be tested " of the present invention refers to that may include containing a kind of mixture of analyte, analyte
But it is not limited to protein, hormone, antibody or antigen.The typical sample to be tested that can be used in method disclosed by the invention includes
Serum and urine.Sample to be tested can be using it is preceding it is as needed using dilution or buffer solution to analyte may be contained
Sample be diluted after solution.For example, in order to avoid HOOK effects, it can use Sample dilution will before upper machine testing
Analyte is detected on detecting instrument again after being diluted, and solution that at this time may be after the dilution containing analyte is equal
It is referred to as sample to be tested.
Term " antibody " of the present invention and " immunoglobulin " are used with most wide meaning, include the antibody of any isotype
Or immunoglobulin, retain the antibody fragment of the specific binding to antigen, including but not limited to Fab, Fv, scFv and Fd piece
Section, chimeric antibody, humanized antibody, single-chain antibody, bispecific antibody and the antigen-binding portion thereof comprising antibody and non-antibody
The fusion protein of albumen.In the case of any need, antibody can further with other parts, such as specific binding pair
Member, such as biotin or Streptavidin (a member in biotin-Streptavidin specific binding pair member) etc. are sewed
It closes.
Term " monoclonal antibody " of the present invention refers to the immunoglobulin secreted by the bone-marrow-derived lymphocyte of monoclonal,
It can be prepared by method known in those skilled in the art.
Term " polyclonal antibody " of the present invention refers to the immune globulin generated by more than one bone-marrow-derived lymphocyte clone
White set, can be prepared by method known in those skilled in the art.
Term " antigen " of the present invention is to refer to stimulation body to generate immune response, and with immune response product can resist
Body and sensitized lymphocyte combine in vivo and in vitro, and the substance of immunological effect occurs.
Term " in conjunction with " of the present invention refers to due to interactions such as example covalent, electrostatic, hydrophobic, ion and/or hydrogen bonds,
Including but not limited to two intermolecular direct joints caused by such as salt bridge and the interaction of water bridge.
Term " specific binding " of the present invention refers to that mutual discrimination between two kinds of substances and selective binding are anti-
Answer, said from stereochemical structure angle be exactly conformation between corresponding reactant correspondence.
Term " biotin " of the present invention is widely present in animal vegetable tissue, there are two cyclic structure on molecule,
Respectively imidazolone ring and thiphene ring, wherein imidazolone ring are the main portions combined with Streptavidin.The biotin of activation
It can be coupled with known almost all creatures macromolecular under the mediation of protein cross agent, including protein, nucleic acid, more
Sugar and lipid etc.；And " Streptavidin " is a kind of protein secreted by streptomycete, molecular weight 65kD." Streptavidin "
Molecule is made of 4 identical peptide chains, wherein every peptide chain can combine a biotin.Therefore each antigen or antibody can be same
When be coupled multiple biotin molecules, to generate " tentacle effect " improve sensitivity for analysis.
Term " donor " of the present invention refers to that can generate after the activation by energy or reactive compound and receptor
The sensitizer of the reactive intermediate of such as singlet oxygen of reaction.Donor can be photoactivation (such as dyestuff and aromatic compound)
Or (such as enzyme, metal salt) of chemical activation.In particular embodiments of the invention, the donor is photosensitizer, described
Photosensitizer can be photosensitizer known in the art, preferably stable with respect to the light and not compound with singlet oxygen effecting reaction,
Its non-limiting example includes sub- disclosed in such as United States Patent (USP) US5709994 (patent document is hereby incorporated by reference)
The compounds such as methyl blue, rose-red, porphyrin, phthalocyanine and chlorophyll and these compounds have 1-50 replacing group
Derivative, the substituent group for so that these compounds with more lipophilicity or with more hydrophily, and/or as connection
To the linking group of specific binding pair member.The example of other photosensitizers well known by persons skilled in the art can also be at this
It is used in invention, such as the content described in United States Patent (USP) US6406913, which is hereby expressly incorporated by reference.At this
It invents in other specific embodiments, the donor is other sensitizers of chemical activation, and non-limiting example is certain
Compound, their catalyzing hydrogen peroxides are converted into singlet oxygen and water.The example of some other donor includes：1,4- dicarboxyl second
Base-Isosorbide-5-Nitrae-naphthalene endoperoxides object, 9,10- diphenylanthrancene -9,10- endoperoxides objects etc. heat these compounds or these changes
Conjunction object, which directly absorbs light, can discharge singlet oxygen.
Term " receptor " of the present invention is to refer to react the substance that can generate detectable signal with singlet oxygen.For
Body is by energy or reactive compound induced activation and the singlet oxygen for discharging upper state, and the singlet oxygen of the upper state is by low coverage
From receptor capture, to transmit energy to activate the receptor.In some specific embodiments of the present invention, the receptor is
Such substance, experience and the chemical reaction of singlet oxygen are to form unstable metastable state intermediate, in the metastable state
Mesosome can decompose, and subsequently or simultaneously shine.The exemplary of these substances includes but not limited to：Enol ether, enamine, 9- alkylidenes
Xanthans, 9- alkylidene-N- alkyl Acridane, aromatic ethylene ether, bicyclic ethylene oxide, thioxene, armaticity imidazoles or lucigenin.
In other specific embodiments of the present invention, the receptor is can to react that be formed ketone can be resolved into singlet oxygen
Or the hydroperoxides of carboxylic acid derivates or the olefines of dioxy cyclobutane；The stabilization dioxy ring that can be decomposed by the effect of light
Butane；It can be reacted with singlet oxygen to form the acetylene class of diones；Azo-compound or azo carbonyl compound can be formed
The hydrazone class or hydrazides of object, such as luminol；With the aromatic compounds that can form endoperoxides species.It can be according to the disclosure
Specific, the non-limiting examples of the receptor utilized with claimed invention are recorded in U.S. Patent number US5340716, and (this is special
Sharp document is hereby incorporated by reference).In other specific embodiments of the invention, the receptor includes olefin(e) compound and gold
The case where category chelate is non-particlized and solvable in water-bearing media, this receptor can be found in patent PCT/
US2010/025433 (patent document is hereby incorporated by reference).
In the present invention, the donor can be coated on to be formed on matrix by functional group to be filled with photosensitive chemical combination
The high molecular particle of object can generate singlet oxygen under light excitation, and donor is referred to as photosensitive microballoon or photosensitive micro- at this time
Grain, including the solution of this photosensitive microballoon or photosensitive particulate is properly termed as photosensitive liquid or general liquid；And/or the receptor can be with
It is to be coated on matrix to form the high molecular particle filled with luminophor and lanthanide series by functional group, at this time may be used
To be known as shine microballoon or luminous particle.In this application, system is excited based on the luminescent substance for being coated on matrix surface through light
With energy transmission induced luminescence signal, energy transmission is combined dependent on Ag-Ab causes photosensitive microballoon and luminous microballoon mutual
It is close and realize.There is no need to separation processes.The diameter smaller of nanoparticle, suspendability is stronger, while using three-level
Amplify luminescent system, thus there is higher sensitivity for analysis；Entire detection process is not necessarily to detach binding marker without cleaning
And binding label, therefore the reaction time is shorter；Probe material (emulsion and luminous agent) marks on matrix, rather than marks
In biomolecule, the activity of biomolecule is not influenced, meanwhile, because there are larger specific surface areas for matrix, therefore its surface
On can be coated with more probe materials and biomolecule, cause it in the effective concentration of reagent and sensitivity and detection background etc. side
The performance in face can be more excellent.
" matrix " of the present invention is microballoon or particle known in those skilled in the art, can be any size
, it can be organic or inorganic, can be inflatable or nondistensible, can be porous or non-porous
, with any density, but preferably have and the close density of water, can preferably float in water, and by transparent, partially transparent
Or opaque material is constituted.Described matrix can be with or without charge, when with charge, preferably negative electrical charge.The base
Body can be that solid (such as polymer, metal, glass, organic and inorganic matter such as mineral, salt and diatom), small oil droplet are (such as hydrocarbon
Compound, fluorocarbon, silicic fluid), vesica (such as phosphatide such as synthesis or natural such as cell and organelle
Official).Matrix can be latex particle or other particles, lipid bilayer such as liposome, phosphatide containing organic or inorganic polymer
Vesica, small oil droplet, silicon particle, metal-sol, cell and crystallite dyestuff.Matrix usually has multifunctionality, or can pass through
Special or non-specific covalently or non-covalently interaction and be attached on donor or receptor.Be available there are many functional group or
Person is merged in.Typical functional group includes carboxylic acid, aldehyde radical (such as acetaldehyde), amino, cyano, vinyl, hydroxyl, sulfydryl
Deng.A unrestricted example for being suitable for the invention matrix is carboxy-modified latex particle.This matrix it is detailed
Situation can be found in United States Patent (USP) US5709994 and US5780646 (this two patents document is hereby incorporated by reference).
Term " epitope " of the present invention is any egg for referring to specific binding immunoglobulin or T cell receptor
White determinant.In some specific embodiments of the present invention, epitope is that antigenic surface can be by the region of antibody specificity set.
Epitopic determinants usually may include the chemically active surface group of molecule, such as, but not limited to：Amino acid, carbohydrate side chain, phosphinylidyne
Base and/or sulfonyl.In some other specific embodiment of the present invention, epitope can specific specific three-valued structures feature and
Specific charge feature.
Term " homogeneous immunological detection reagent suit " of the present invention refers to the whole that homogeneous immune detection must use
The combination of reagent or medicament.
II, specific embodiments
The testing principle of the present invention is as shown in Figure 1：It is anti-in receptor surface coating mouse using homogeneous luminescent method of immunity
People's FSH monoclonal antibodies, obtain the receptor combined with antibody；By the anti-human FSH labeling of monoclonal antibodies biotin of another plant of mouse, system
The standby antibody combined with biotin；Meanwhile in donor pan coating Streptavidin, obtaining the confession combined with Streptavidin
Body.
When detecting that there are FSH to be measured, FSH to be combined simultaneously with the specific antibody of receptor surface and with biotin in system
Specific antibody combine, formed double-antibody sandwich compound；Two kinds of antibody excess at this time, there are the antibody of unbound state point
Son.It is added the donor that is combined with Streptavidin, Streptavidin is combined with biotin molecule (including compound or to dissociate biological
Plain antibody), but the biotin in compound (FG-Ab-FSH-Ab-Bio) is only combined, the distance of receptor and donor could be drawn
Closely, under the excitation of energy or reactive compound, donor discharges singlet oxygen, and chemistry is generated after touching receptor in the solution
It shines, fluorescence is generated to further excite the fluorophor on receptor to generate Cascaded amplification reaction.It is free with antibody knot
The receptor of conjunction cannot obtain energy far from donor, generate optical signal.Therefore, light signal strength is in the FSH contents in sample
Direct proportion function relationship.Mathematical function relationship, Bian Keji are established using known FSH concentration calibrations product and corresponding light signal strength
Calculate the concentration level of unknown FSH samples to be measured.
In certain specific embodiments of the invention, prepared by the method for the invention detects the homogeneous of follicular stimulating hormone
Immunologic function test reagent is set with, including：
(1) FSH calibration objects serial dilutions：6 concentration are 200mIU/ml, 100mIU/ml, 50mIU/ml, 10m respectively
IU/ml、1mIU/ml、0mIU/ml；
(2) the anti-human FSH monoclonal antibody solutions of mouse combined with biotin, a concentration of 2~8 μ g/mL；
(3) the receptor microspheres solution combined with the anti-human FSH monoclonal antibodies of mouse, a concentration of 0.2~0.8mg/mL；
(4) the donor microspheres solution combined with Streptavidin.
The anti-human FSH monoclonal antibodies of mouse are commercial reagents, are available.
The anti-human FSH monoclonal antibodies of mouse should meet following leading indicator：
Specificity：Immunogene uses people source FSH or reorganization FSH, has a good specificity with people FSH, but with people TSH, LH,
HCG cross reacting rates are less than 0.01%；
Affinity (0~200mIU/ml) in detection range shows good combination, and it is bent can to obtain good dose-response
Line；
Potency enzyme-linked immunosorbent assay (ELISA) is more than 1/15000；
Purity protein A affinity chromatography is pure, and purity is more than 99%.
The luminous microballoon (receptor), photosensitive microballoon (donor) are commercial reagents.
Microballoon shine for being combined with the anti-human FSH monoclonal antibodies of mouse.The microballoon that shines is modified with aldehyde radical, passes through aldehyde radical
It is connect with antibody molecule, forms the luminous microballoon combined with antibody.The microballoon that shines is interior to contain luminophor and lanthanide series
The chelate of object is closed, luminophor is the derivative of thioxene, and lanthanide compound can be Eu (TTA) 3/TOPO
Or Eu (TTA) 3/Phne.Receptor microballoon is purchased from platinum Elmer Co., Ltd, article No. 6762001.
Photosensitive microballoon has included Photoactive compounds phthalein mountain valley with clumps of trees and bamboo dyestuff (luminol class chemiluminescent substance), while in photosensitive microballoon
It is combined containing active aldehyde, and with Streptavidin.Photosensitive microballoon is purchased from platinum Elmer Co., Ltd, article No.
6760002S。
Further include opaque microwell plate, such as 96 hole reaction plate of homogeneous luminescent in the reagent set.
III, embodiments
To keep the present invention easier to understand, below in conjunction with embodiment, present invention be described in more detail, these realities
Apply example only serve it is illustrative, it is not limited to application range of the invention.If the raw material used in the present invention or component nothing
Specified otherwise can be made by commercial sources or conventional method.
Embodiment 1：Prepare the homogeneous immunological detection reagent suit of detection follicular stimulating hormone
(1) preparation of the receptor combined with antibody：
Receptor as the present embodiment is to be coated on to be formed on latex particle containing aldehyde radical (- CHO) to be filled with dimethyl thiophene
The particle (luminous microballoon) of the derivative of pheno and the chelate of lanthanide series Eu.Receptor is connect by aldehyde radical with antibody molecule.
Biological raw material：FSH monoclonal antibodies
Preparation process：It takes 2mg to shine microspheres solution, the carbonate buffer solution (CB) of 0.05M pH 9.6 is used in combination to be diluted
At 5mg/ml；Take FSH monoclonal antibodies 0.02mg be transferred to dilution after luminous microspheres solution in, mix well, 4 DEG C overnight；
The BSA solution 20uL that 10mg/ml is diluted to the CB buffer solutions of 0.05M pH 9.6 are added, room temperature rotates 2h；Fully washing
Microballoon is finally diluted to 0.5mg/ml with the Tris-HCl solution of 8.0 0.1M of pH, obtains reagent R1 by microballoon.
(2) preparation process of the antibody combined with biotin labeling
Biological raw material：Activated biotin and FSH monoclonal antibodies
Preparation process：FSH monoclonal antibodies 0.5mg is taken to be transferred in 14KD bag filters, with label buffer solution (0.1M
NaHCO3) dialysis, 2h/ times, change liquid 1 time；The biotin solution 10ul of 5mg/ml, rapid mixing, supplemental markers buffer solution is added
To 500 μ l, 2-8 DEG C of mixing overnight, label ratio is 1:30 (antibody:Biotin-molar ratio)；The Bio-Ab that label is completed turns
14KD bag filters are moved to, is dialysed with elution buffer (0.1MTris-HCl), 2h/ times, changes liquid 1 time；With 8.0 0.1M of pH
Tris-HCl solution is diluted to 5 μ g/ml, obtains reagent R2.
(3) preparation of the donor combined with Streptavidin
A, photosensitive microballoon (donor) suspension processing：Draw a certain amount of photosensitive microballoon in high speed freezing centrifuge from
The heart discards supernatant, and a certain amount of MES buffer solutions are added, and ultrasound suspends again to particle on ultrasonic cell disintegration instrument, and it is slow that MES is added
Fliud flushing adjusts photosensitive microballoon concentration to 100mg/ml.
B, solution of streptavidin is prepared：A certain amount of Streptavidin is weighed, adds MES buffer solutions to 8mg/ml.
C, it mixes：Photosensitive microballoon (donor) suspension handled well, the solution of streptavidin of 8mg/ml and MES is slow
Fliud flushing, with 2:5:1 volume ratio is mixed, and rapid mixing obtains reaction solution.
D, it reacts：The NaBH of MES buffers 25mg/ml3CN solution, according to reaction solution 1:25 volume ratio adds
Enter, rapid mixing, 37 DEG C of revolving reactions 48 hours.
E, it closes：The Gly solution of MES buffers 75mg/ml and the NaBH of 25mg/ml3CN solution, according to it is anti-
Answer liquid 2:1:10 volume ratio is added in above-mentioned solution, mixing, 37 DEG C of revolving reactions 2 hours.The BSA for adding 200mg/ml is molten
Liquid (MES buffer solutions) is 5 with reaction solution volume ratio:8, rapid mixing, 37 DEG C of revolving reactions 16 hours.
F, it cleans：MES buffer solutions are added into completely reacted solution, supernatant is abandoned in high speed freezing centrifuge centrifugation, is added new
Fresh MES buffer solutions ultrasonic method suspends again, centrifuges again, so cleaning 3 times, is finally suspended with photo-sensitire reagent buffer solution,
Obtain third composition.Wherein, in third composition component c a concentration of 100 μ g/mL, used as general liquid, obtain reagent
R3。
(4) preparation process of FSH calibration objects serial dilutions
FSH sterlings are taken, are configured to respectively with 7.4 phosphate buffered saline solutions of 0.1M pH containing 20% inactivation calf serum
0.5ml 0,1,10,50,100,200mIU/mL calibration object serial dilutions.
Embodiment 2：Follicular stimulating hormone in the reagent set detection sample prepared using the method for the invention.
Used reagent set is the homogeneous immunological detection reagent set for the follicular stimulating hormone that embodiment 1 is prepared.Inspection
Survey process is automatically finished and exports testing result, specific steps by automatic light-induced chemiluminescent analysis system：
1) 25 μ l samples to be tested or calibration object, quality-control product are separately added into reacting hole；
2) 25 μ LR1 and 25 μ l R1 are sequentially added in reacting hole；
3) it incubates 15 minutes for 37 DEG C；
4) 175 μ l R3 are added；
5) it incubates 15 minutes for 37 DEG C；
6) signal value of laser irradiation micropore and calculating per hole；Standard curve is drawn according to the signal value of calibration object, and is led to
Standard curve is crossed, the concentration of follicular stimulating hormone in sample to be tested is calculated.Measurement result difference is as shown in Table 1, 2 and 3, measures knot
Fruit and the correlation of Beckman definite value are as shown in Figure 2.
Table 1：The measurement result of sample 1
Table 2：The measurement result of sample 2
Table 3：The measurement result of sample 3
From table 1-3 and Fig. 2 it is found that the follicular stimulating hormone concentration and the Beckman measured value that are measured using the method for the invention
Correlation r=0.9955, correlation is good, illustrates accurately detect that ovarian follicle stimulates in sample using the method for the invention
The content of element.
Embodiment 3：The detection precision of reagent set prepared by the method for the invention
Precision is to weigh the important indicator of reagent within-run and between-run analysis, is that the important of commercialized product validity is intended in evaluation
Foundation generally includes withinrun precision and betweenrun precision.
Withinrun precision appraisal procedure：With low (L), in (M), high (H) value sample, the product of 3 batches is carried out independent
Analysis, each batch replication 10 times (being measured using method described in embodiment 2), 10 measurement results of calculating
Average value (x) and standard deviation (SD) calculate the coefficient of variation (CV), the results are shown in Table 4 according to formula CV=SD/x × 100%.
Betweenrun precision appraisal procedure：With low (L), in (M), high (H) value sample, the product of 3 batches is carried out independent
Analysis, each batch replication 10 times (being measured using the method that embodiment 2 describes), 30 measurement results of calculating are put down
Mean value (x) and standard deviation (SD) calculate the coefficient of variation (CV), the results are shown in Table 5 according to formula CV=SD/x × 100%.
Table 4：The withinrun precision of reagent set prepared by the method for the invention
Table 5：The betweenrun precision of reagent set prepared by the method for the invention
From table 4 and table 5 it is found that the withinrun precision and betweenrun precision of reagent set prepared by the method for the invention are equal
<5%, illustrate that the reproducible, random error when reagent set prepared using the method for the invention is detected is small.
Embodiment 4：The accuracy in detection of reagent set prepared by the method for the invention
Accuracy is the degree that is consistent of measured value and actual value, and reaction reagent is set with the size of detection error.
Evaluation of accuracy：By 2 samples (sample 1 and sample 2) of the level containing different FSH, with calibration object dilution
Multipoint dilution is carried out, the concentration of sample after dilution is measured using method described in embodiment 2, and dilute according to dilution ratio meter
The rate of recovery is calculated, as a result as shown in table 6 and table 7.
Table 7：The accuracy in detection to sample 1 of reagent set prepared by the method for the invention
Table 8：The accuracy in detection to sample 2 of reagent set prepared by the method for the invention
It is measured after from table 7 and table 8 it is found that carrying out multipoint dilution with the FSH samples of 2 different levels, the rate of recovery is equal
In 90%~110% range, illustrate the detection for the reagent set that measured value is close with actual value, prepared by the method for the invention
Error is small.
It should be noted that embodiment described above is only used for explaining the present invention, do not constitute to any of the present invention
Limitation.By referring to exemplary embodiments, invention has been described, it should be appreciated that word used in it is descriptive
With explanatory vocabulary, rather than limited vocabulary.The present invention can be made within the scope of the claims by regulation
Modification, and the present invention is revised in without departing substantially from scope and spirit of the present invention.Although the present invention described in it relates to
And specific method, material and embodiment, it is not intended that the present invention is limited to particular case disclosed in it, on the contrary, this hair
It is bright to can be extended to other all methods and applications with the same function.
Claims (16)
1. a kind of method for the homogeneous immunological detection reagent suit preparing detection follicular stimulating hormone comprising：
First chamber is prepared, the first chamber includes component a and the first buffer solution, and wherein component a is and the first knot
Close the receptor that generation detectable signal can be reacted with singlet oxygen that unit combines；First combining unit can be with ovarian follicle
First epitope specificity of stimulin combines；
Second chamber is prepared, the second chamber includes component b and the second buffer solution, and wherein component b is and the first mark
Remember the second combining unit that object combines；Second combining unit can be combined with the second epitope specificity of follicular stimulating hormone,
And second epitope and the first epitope non-overlapping；
Third composition is prepared, the third composition includes component c and third buffer solution, wherein the component c is and the
The donor that singlet oxygen can be generated in excited state that the specific junction mixture of one marker combines.
2. according to the method described in claim 1, it is characterized in that, first combining unit and the second combining unit is selected from
Polyclonal antibody, monoclonal antibody, antibody binding fragment, artificial antibody, modification antibody, be preferably selected from monoclonal antibody and/
Or polyclonal antibody.
3. method according to claim 1 or 2, which is characterized in that a concentration of the 0.2 of component a in the first chamber
~0.8mg/mL；Preferably, in the first chamber component a a concentration of 0.4~0.6mg/mL；And/or
A concentration of 2~8 μ g/mL of component b in the second chamber；Preferably, in the second chamber component b concentration
For 4~6 μ g/mL；And/or
A concentration of 50~150 μ g/mL of component c in the third composition；Preferably, component c in the third composition
A concentration of 80~120 μ g/mL.
4. according to the method described in any one of claim 1-3, which is characterized in that the method further includes preparing ovarian follicle thorn
Hormone calibration object serial dilutions.
5. according to the method described in claim 4, it is characterized in that, the concentration of the follicular stimulating hormone calibration object serial dilutions
For 0~200mIU/mL；Preferably, the concentration of the follicular stimulating hormone calibration object serial dilutions is respectively 0mIU/mL, 1mIU/
ML, 10mIU/mL, 50mIU/mL, 100mIU/mL and 200mIU/mL.
6. according to the method described in any one of claim 1-5, which is characterized in that the preparation method of the first chamber
Including：
Receptor is diluted to 3~8mg/mL using carbonate buffer solution, obtains receptor solution by step S1；
The first combining unit is added in step S2 in receptor solution, added after static and be diluted to 9 with carbonate buffer solution~
The BSA solution of 10mg/mL obtains the receptor solution combined with the first combining unit；
Step S3 detaches the receptor combined with the first combining unit in the receptor solution combined with the first combining unit, and adds
Enter the first buffer solution, obtains first chamber.
7. according to the method described in any one of claim 1-6, which is characterized in that the preparation method of the second chamber
Including：
Second combining unit is placed in bag filter and is dialysed using label buffer solution, the is added after dialysis by step T1
One marker solution, and elution buffer is filled into, it is static, obtain the second combining unit combined with the first marker；
The second combining unit combined with the first marker is placed in bag filter and is carried out thoroughly using elution buffer by step T2
The second buffer solution is added in analysis after dialysis, obtain second chamber solution.
8. the method according to the description of claim 7 is characterized in that the NaHCO that the label buffer solution is 0.09~0.1M3It is molten
Liquid；The elution buffer is the Tris-HCl solution of 0.09~0.1M；And/or
The molar ratio of second combining unit and the first marker is 1:(20~40).
9. according to the method described in any one of claim 1-8, which is characterized in that first buffer solution and second delays
Rush the Tris-HCl solution that solution is 0.09~0.10M that pH value is 7.0~8.5.
10. according to the method described in any one of claim 1-9, which is characterized in that the receptor includes olefin(e) compound
And metallo-chelate, it is non-particulate forms, and solvable in water-bearing media；And/or the receptor is filled with light emitting compound
The high molecular particle of object and lanthanide series；
The donor is photoactivation or chemical activation sensitizer, is non-particulate forms, and solvable in water-bearing media；
And/or the donor is the high molecular particle filled with Photoactive compounds.
11. a kind of homogeneous immune detection examination for the detection follicular stimulating hormone for wanting any one of 1-10 the methods to prepare such as right
Agent is set with.
12. a kind of homogeneous immunization method detecting follicular stimulating hormone using reagent set as claimed in claim 11, the side
Method detects for light-induced chemiluminescent.
13. according to the method for claim 12, which is characterized in that the method includes：
Sample to be tested is mixed with first chamber and second chamber, obtains the first mixture by step R1；
First mixture is mixed with third composition, obtains the second mixture by step R2；
Step R3 excites the donor in the second mixture to generate singlet oxygen, the receptor using energy or reactive compound
It can be reacted with the singlet oxygen received and generate detectable chemiluminescence signal；
The presence of the chemiluminescence signal obtained in step R4, detecting step R3 and/or intensity, to judge to survey in sample to be tested
With the presence or absence of the content of follicular stimulating hormone and/or determining follicular stimulating hormone.
14. according to the method for claim 13, which is characterized in that the method further includes：It is calibrated using follicular stimulating hormone
Product serial dilutions make the step of standard curve of chemiluminescence signal-follicular stimulating hormone concentration；The standard curve is used for
Determine the content of follicular stimulating hormone in sample to be tested.
15. according to the method for claim 13, which is characterized in that the described method comprises the following steps：
A. the sample to be tested of 10~30 μ L is added in reacting hole；
B. second chamber described in first chamber and 10~30 μ L described in 10~30 μ L is sequentially added in reacting hole, wherein the
A concentration of 0.2~0.8mg/mL of component a in one composition, a concentration of 2~8 μ g/mL of the component b in second chamber；
C.35~45 DEG C incubation 10~30 minutes；
D. 100~250 μ L of the third composition are added into reacting hole；Component c's wherein in third composition is a concentration of
50~150 μ g/mL；
E.35~45 DEG C incubation 5~20 minutes；
F. it is the laser irradiation reacting hole of 680nm to utilize wavelength, and excited donor generates singlet oxygen, receptor and the single line touched
The reaction of state oxygen produces the transmitting light of 612nm；
G. the photon amount for detecting transmitting light in each reacting hole, the concentration of follicular stimulating hormone is calculated according to standard curve.
16. a kind of reagent set as claimed in claim 11 is being prepared for detecting follicular stimulating hormone horizontal abnormality associated disease
Kit in application；Preferably, the illness includes primary amenorrhea, primary hypogona dism, early stage anterior pituitary
Hyperfunction, seminoma of testis, estrogen or progesterone treatment secondary hypogonadism and Skien syndrome late period hang down
Body function is low.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810142754.9A CN108490194A (en) | 2018-02-11 | 2018-02-11 | Detect the homogeneous immunological detection reagent suit and its preparation method and application of follicular stimulating hormone |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810142754.9A CN108490194A (en) | 2018-02-11 | 2018-02-11 | Detect the homogeneous immunological detection reagent suit and its preparation method and application of follicular stimulating hormone |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108490194A true CN108490194A (en) | 2018-09-04 |
Family
ID=63340347
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810142754.9A Pending CN108490194A (en) | 2018-02-11 | 2018-02-11 | Detect the homogeneous immunological detection reagent suit and its preparation method and application of follicular stimulating hormone |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108490194A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1745300A (en) * | 2002-06-12 | 2006-03-08 | 塞诺菲-安万特德国有限公司 | HTS-capable method and testing system for determining the interaction between a C-reactive protein and constituents that bind to a C-reactive protein |
| CN101377490A (en) * | 2007-08-30 | 2009-03-04 | 北京科美东雅生物技术有限公司 | Magnetic microparticle separating chemiluminescence immune analysis determination reagent kit for detecting related sign object and preparing method thereof |
| CN101769933A (en) * | 2008-12-30 | 2010-07-07 | 博阳生物科技（上海）有限公司 | Detection micro particle of thyroid stimulating hormone and preparation and application thereof |
| CN102348980A (en) * | 2009-03-12 | 2012-02-08 | 西门子医疗保健诊断公司 | Immunoassays employing non-particulate chemiluminescent reagent |
| CN102798725A (en) * | 2012-08-13 | 2012-11-28 | 沃克(天津)生物科技有限公司 | Diagnostic kit for determination of serum total IgE, preparation method and application method |
-
2018
- 2018-02-11 CN CN201810142754.9A patent/CN108490194A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1745300A (en) * | 2002-06-12 | 2006-03-08 | 塞诺菲-安万特德国有限公司 | HTS-capable method and testing system for determining the interaction between a C-reactive protein and constituents that bind to a C-reactive protein |
| CN101377490A (en) * | 2007-08-30 | 2009-03-04 | 北京科美东雅生物技术有限公司 | Magnetic microparticle separating chemiluminescence immune analysis determination reagent kit for detecting related sign object and preparing method thereof |
| CN101769933A (en) * | 2008-12-30 | 2010-07-07 | 博阳生物科技（上海）有限公司 | Detection micro particle of thyroid stimulating hormone and preparation and application thereof |
| CN102348980A (en) * | 2009-03-12 | 2012-02-08 | 西门子医疗保健诊断公司 | Immunoassays employing non-particulate chemiluminescent reagent |
| CN102798725A (en) * | 2012-08-13 | 2012-11-28 | 沃克(天津)生物科技有限公司 | Diagnostic kit for determination of serum total IgE, preparation method and application method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108051585A (en) | A kind of homogeneous immunological detection reagent box, detection method and its application | |
| CN108169497A (en) | Human prolactin detection kit and preparation method and application | |
| CN109709317B (en) | Homogeneous phase immunoassay kit without matrix effect and analysis method and application thereof | |
| CN110161230A (en) | Quickly homogeneous immune reagent kit, preparation method, detection method and the device of detection Procalcitonin | |
| WO2007121668A1 (en) | Direct chemiluminescent reagent for magnetic separation and immunoassay method using thereof | |
| CN108445239A (en) | Detect the homogeneous immunological detection reagent suit and its preparation method and application of β human chorionic gonadotrophins | |
| CN108445223A (en) | Detect homogeneous immunological detection reagent box and its application of the anti-Carp antibody of target | |
| CN109725153A (en) | Homogeneous phase immunoassay method and application thereof | |
| CN109211867A (en) | A kind of micro-fluidic fluorescence immunoassay chip of rapid quantitative detection BNP | |
| CN106918708A (en) | A kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin | |
| CN113125696B (en) | Estradiol homogeneous chemiluminescence detection kit and application thereof | |
| CN108445216A (en) | A kind of anti-Miao Le Shi pipes hormone determination kit of people and the preparation method and application thereof | |
| CN110161247A (en) | Detect homogeneous immunological detection reagent box and its application of cyclic citrullinated peptid | |
| TW201802472A (en) | Anti-human hemoglobin monoclonal antibody or antibody kit, insoluble carrier particle to which anti-human hemoglobin monoclonal antibody is immobilized, and measurement reagent and measurement method using same | |
| CN110161250A (en) | A kind of homogeneous human cardiac troponin I quick detection kit, system, detection method and application | |
| CN106460056A (en) | Novel method for detecting detection object in sample, and detection kit using same | |
| CN113125731A (en) | Competitive homogeneous phase chemiluminescence assay kit and application thereof | |
| CN108490194A (en) | Detect the homogeneous immunological detection reagent suit and its preparation method and application of follicular stimulating hormone | |
| CN114354919A (en) | Homogeneous immunoassay kit for detecting 14-3-3eta protein and application thereof | |
| CN108918888A (en) | It is a kind of detect Endostatin latex enhancing immune than turbid kit and its application | |
| CN113125697B (en) | Testosterone homogeneous phase chemiluminescence detection kit and application thereof | |
| CN108445238A (en) | Detect the homogeneous immunological detection reagent suit and its preparation method and application of interstitialcellstimulating hormone (ICSH) | |
| CN110514648A (en) | A kind of chemiluminescence analysis POCT detection device and its application | |
| CN111217910A (en) | Monoclonal antibody pair and application thereof in detecting myeloperoxidase protein | |
| CN111505302A (en) | Homogeneous immunoassay kit for rapidly detecting N-terminal B-type brain natriuretic peptide, preparation method, detection method and device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 100094 Beijing Haidian District Yongfeng Base Fengxianzhong Road 7 Beike Modern Manufacturing Park Incubation Building Applicant after: Kemei Diagnostic Technology Co., Ltd Address before: 100094, two floor, B block, North Branch modern manufacturing park, 7 Fengxian Middle Road, Yongfeng base, Haidian District, Beijing. Applicant before: Beijing Kemei Biological Technology Co., Ltd. |
|
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Xu Jingxin Inventor after: Qiang Zhonghua Inventor after: Liu Yuhui Inventor after: Li Lin Inventor before: Xu Jingxin Inventor before: Qiang Zhonghua Inventor before: Li Huiqiang Inventor before: Liu Yuhui Inventor before: Li Lin |
|
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180904 |