CN108828207A - A kind of excellent effect cadmium chelating type immune complex and preparation method thereof - Google Patents
A kind of excellent effect cadmium chelating type immune complex and preparation method thereof Download PDFInfo
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- CN108828207A CN108828207A CN201810594250.0A CN201810594250A CN108828207A CN 108828207 A CN108828207 A CN 108828207A CN 201810594250 A CN201810594250 A CN 201810594250A CN 108828207 A CN108828207 A CN 108828207A
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Abstract
The invention discloses a kind of excellent effect cadmium chelating type immune complex, which is following one kind：Cadmium ion is incorporated into the compound of immune complex formation；Or cadmium ion be incorporated into after carrier protein with and the antibody that specifically binds of the carrier protein be formed by compound；Or cadmium ion is incorporated into the compound formed in conjunction with carrier protein after immunoglobulin.The invention also discloses a kind of preparation methods of excellent effect cadmium chelating type immune complex, include the following steps：S1：Prepare chelating agent solution, S2：Formulation vehicle protein solution, S3：It is stirred overnight, S4：Dialysis treatment, S5：Cadmium ion, S6 is added：Devil liquor recovery processing；S7：It is specifically bound.The method of the present invention scope of application is wider, can be with save the cost, and improves dialysis rate, can shorten manufacturing cycle, also have the characteristics that energy conservation and environmental protection, avoids chemical contamination, and environment protecting is good.
Description
Technical field
The present invention relates to a kind of chelate field, specially a kind of excellent effect cadmium chelating type immune complex and its preparation
Method.
Background technique
Chelate Mineral Floating Process, hydrometallurgy, metallic element extraction with separate, the catalyzing and synthesizing of substance, water
Softening, electroplating technology, medical industry, dyeing course etc. in be all widely used；Existing cadmium chelating type immune complex
Preparation method it is incomplete, the scope of application is ignored enough wide, it has not been convenient to the preparation process of object is completed in small-size laboratory,
And number of dialysing is excessive, can extend the manufacturing cycle of object to a certain extent, and there are no give up to the ion etc. in waste liquid
Object, which is done, clearly to be handled, and the wastes such as ion in waste liquid, which do not handle rear discharge up to standard, will cause chemical contamination.
Summary of the invention
The purpose of the present invention is to provide a kind of excellent effect cadmium chelating type immune complexs and preparation method thereof, to solve
The problems mentioned above in the background art.
To achieve the above object, the present invention provides the following technical solutions：A kind of excellent effect cadmium chelating type immune complex,
The cadmium chelating type immune complex is following one kind:
Cadmium ion is incorporated into the compound of immune complex formation；
Or cadmium ion be incorporated into after carrier protein with and the antibody that specifically binds of the carrier protein be formed by compound；
Or cadmium ion is incorporated into the compound formed in conjunction with carrier protein after immunoglobulin.
Preferably, zinc fingers, sulfydryl, cysteine are at least contained in the structure of the immune complex or carrier protein
One of residue, cadmium ion are mutually tied with immune complex or carrier protein with zinc fingers or sulfydryl or cysteine residues
It closes.
Preferably, the carrier protein is antibody protein, lipoprotein, hemoglobin, nuclear antigen, foreign sera antigen, disease
Poison, bacterium, protozoon, worm or immunoglobulin one of.
Preferably, it is specifically bound again with energy and carrier protein after cadmium ion passes through chelating agent in conjunction with carrier protein anti-
Body combines, and the chelating agent is one of ITCBE, EDTA, polyphosphate, amino carboxylic acid, 1,3- diketone, hydroxycarboxylic acid.
A kind of preparation method of cadmium chelating type immune complex as described in claim 1, includes the following steps：
S1：First chelating agent solution is prepared and is completed, then chelating agent is dissolved in solvent, chelating agent solution can be obtained；
S2：Appropriate carrier protein is taken, carrier protein is added in borate buffer solution, it is molten to can be prepared by carrier protein
Liquid；
S3：Chelating agent solution obtained is added in carrier protein solution, is placed in water bath with thermostatic control and stirs evenly,
Water bath with thermostatic control 4-30h, can be obtained mixed liquor；
S4：It fetches bag filter and EDTA-NaHCO is added in bag filter3Solution boils, and bag filter is fixed on water tank
Middle part, and enough ddH are added in water tank2O, the ddH2The volume ratio of solution is 85-90 in the volume and bag filter of O:1, weight
Multiple the step 2-4 times, the mixing in S3 is fitted into bag filter, bag filter is placed in constant temperature water tank, added in constant temperature water tank
Enter enough ddH2O, the ddH2The volume ratio of solution is 85-90 in the volume and bag filter of O:1, after dialysed overnight, it will dialyse
Liquid in bag is collected.
S5：PH to 7.0 is adjusted dilute hydrochloric acid solution is added from the liquid collected in bag filter, it is molten to be slowly dropped into cadmium ion
Liquid, when instillation, do not stop to shake, and instill and complete to be placed in water bath with thermostatic control and stir evenly, water bath with thermostatic control 4-30h, then repeatedly S4
Once, liquid is collected, cadmium chelating type antigen is obtained.
S6：The waste liquid collected in solution and S5 in S4 outside bag filter is mixed, is added into mixed solution
Enter the cellulosic material being modified to be adsorbed, and repeat to add the above-mentioned cellulosic material being modified, reuses Atomic absorption
Method carries out quantitative analysis to processed solution, and when cadmium ion enrichment degree reaches 0.1mol/ml in solution, recycling cadmium ion is again
It utilizes.
S7：The antibody that can be specifically bound with carrier protein is added in above-mentioned cadmium chelating type antigen, obtains cadmium after reaction
Chelating type immune complex.
The present invention also provides above-mentioned cadmium chelating type immune complexs to detect cadmium chelating type CIC ELISA in preparation
Reagent or enzyme linked immunological kit in application.
The present invention also provides a kind of above-mentioned enzyme linked immunological kit for detecting cadmium chelating type CIC ELISA, the examinations
Agent box includes the standard items of above-mentioned cadmium chelating type immune complex.
Preferably, mentioned reagent box further includes at least one following reagent：Albumen containing capture antigen antibody complex
Coating buffer, the coating buffer of antibody containing capture cadmium, enzyme labelled antibody.
A kind of method of quantitative detection cadmium chelating type CIC ELISA, it is immune with the above-mentioned cadmium chelating type of known content
Compound is detected as standard items using a pair of sample of following methods：Enzyme-linked immunization, enzyme linked immunological and Atomic absorption
Spectrum combined techniques, enzyme linked immunological and inductivity coupled plasma mass spectrometry combined techniques, purification immune complex and atomic absorption spectrum
Combined techniques, purification immune complex and inductivity coupled plasma mass spectrometry combined techniques, electrophoresis and enzyme linked immunological or atomic absorption light
Spectrum or inductivity coupled plasma mass spectrometry combined techniques.
Compared with prior art, the beneficial effects of the invention are as follows：The method of the present invention scope of application is wider, can save into
This, and increase the differential water pressures inside and outside bag filter, dialysis rate is improved, manufacturing cycle can be shortened, also to the ion in waste liquid
Equal wastes are recycled, and have the characteristics that energy conservation and environmental protection, avoid chemical contamination, therefore the present invention has biggish city
Field competitiveness.
Detailed description of the invention
Fig. 1 is flow diagram of the invention.
Specific embodiment
The technical scheme in the embodiments of the invention will be clearly and completely described below, it is clear that described implementation
Example is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, this field is common
Technical staff's every other embodiment obtained without making creative work belongs to the model that the present invention protects
It encloses.
The present invention provides a kind of technical solution：A kind of excellent effect cadmium chelating type immune complex, the cadmium chelating type are immune
Compound is following one kind:
Cadmium ion is incorporated into the compound of immune complex formation；
Or cadmium ion be incorporated into after carrier protein with and the antibody that specifically binds of the carrier protein be formed by compound；
Or cadmium ion is incorporated into the compound formed in conjunction with carrier protein after immunoglobulin.
Further, at least residual containing zinc fingers, sulfydryl, cysteine in the structure of immune complex or carrier protein
One of base, cadmium ion are combined with immune complex or carrier protein with zinc fingers or sulfydryl or cysteine residues.
Further, at least residual containing zinc fingers, sulfydryl, cysteine in the structure of immune complex or carrier protein
One of base, cadmium ion are combined with immune complex or carrier protein with zinc fingers or sulfydryl or cysteine residues.
Further, carrier protein be antibody protein, lipoprotein, hemoglobin, nuclear antigen, foreign sera antigen, virus,
Bacterium, protozoon, worm or immunoglobulin one of.
A kind of preparation method of cadmium chelating type immune complex as claimed in claim 1, includes the following steps：
S1：First chelating agent solution is prepared and is completed, then chelating agent is dissolved in solvent, chelating agent solution can be obtained；
S2：Appropriate carrier protein is taken, carrier protein is added in borate buffer solution, it is molten to can be prepared by carrier protein
Liquid；
S3：Chelating agent solution obtained is added in carrier protein solution, is placed in water bath with thermostatic control and stirs evenly,
For 24 hours, mixed liquor can be obtained in water bath with thermostatic control；
S4：It fetches bag filter and EDTA-NaHCO is added in bag filter3Solution boils, and bag filter is fixed on water tank
Middle part, and enough ddH are added in water tank2O, the ddH2The volume ratio of solution is 85 in the volume and bag filter of O:1, it repeats
Mixing in S3 is fitted into bag filter, bag filter is placed in constant temperature water tank by the step 2 time, and foot is added in constant temperature water tank
The ddH of amount2O, the ddH2The volume ratio of solution is 85 in the volume and bag filter of O:1, it, will be in bag filter after dialysed overnight
Liquid is collected.
S5：PH to 7.0 is adjusted dilute hydrochloric acid solution is added from the liquid collected in bag filter, it is molten to be slowly dropped into cadmium ion
Liquid, when instillation, do not stop to shake, and instill and complete to be placed in water bath with thermostatic control and stir evenly, water bath with thermostatic control for 24 hours, then repeatedly S4 mono-
It is secondary, liquid is collected, cadmium chelating type antigen is obtained.
S6：The waste liquid collected in solution and S5 in S4 outside bag filter is mixed, is added into mixed solution
Enter the cellulosic material being modified to be adsorbed, and repeat to add the above-mentioned cellulosic material being modified, reuses Atomic absorption
Method carries out quantitative analysis to processed solution, and when cadmium ion enrichment degree reaches 0.1mol/ml in solution, recycling cadmium ion is again
It utilizes.
S7：The antibody that can be specifically bound with carrier protein is added in above-mentioned cadmium chelating type antigen, obtains cadmium after reaction
Chelating type immune complex.
The present invention also provides above-mentioned cadmium chelating type immune complexs to detect cadmium chelating type CIC ELISA in preparation
Reagent or enzyme linked immunological kit in application.
The present invention also provides a kind of above-mentioned enzyme linked immunological kit for detecting cadmium chelating type CIC ELISA, the examinations
Agent box includes the standard items of above-mentioned cadmium chelating type immune complex.
Mentioned reagent box further includes at least one following reagent：The coating of albumen containing capture antigen antibody complex
Liquid, the coating buffer containing the antibody for capturing cadmium, enzyme labelled antibody.
In the present invention, the kit for being able to achieve the object of the invention can be listed following several, and but it is not limited to this.
It is a kind of for detecting the kit of cadmium chelating type CIC ELISA, including：It is multiple containing antigen-antibody can be captured
Close coating buffer, confining liquid, washing buffer, Cadmium resistance antibody, enzyme labelled antibody, substrate, the terminate liquid, dilution buffer of the albumen of object
Liquid, positive control, negative control.
It is a kind of for detecting the kit of cadmium chelating type CIC ELISA, including：It is multiple containing antigen-antibody can be captured
Close coating buffer, confining liquid, the washing buffer, eluent, positive control, negative control of the albumen of object.
It is a kind of for detecting the kit of cadmium chelating type CIC ELISA, including：It is multiple containing antigen-antibody can be captured
Close coating buffer, confining liquid, the washing buffer, eluent, nitric acid, hydrogen peroxide, positive control, negative control of the albumen of object.
It is a kind of for detecting the kit of cadmium chelating type CIC ELISA, including for extracting needed for immune complex
Solution redissolves liquid, the coating buffer containing the antibody that can capture cadmium, confining liquid, washing buffer, enzyme labelled antibody, substrate, termination
Liquid, dilution buffer, standard items, negative control.
It is a kind of for detecting the kit of cadmium chelating type CIC ELISA：Including for extracting needed for immune complex
Solution redissolves liquid, positive control, negative control.
In above-mentioned several kits, coating buffer be capture CIC ELISA albumen, as C1Q, CIF albumen,
Anti-C_3 antibody；Confining liquid is the bovine serum albumin(BSA) or skimmed milk power that mass concentration is 2%-4%；Eluent includes but is not limited to
The Tris-HCl buffer solution of papain；Redissolving liquid is to include but is not limited to；Glue bed medium includes but is not limited to agarose
Gel, polyacrylamide gel；It include but is not limited to PEG solution, boric acid salt buffer for extracting solution needed for immune complex
Liquid；Substrate includes but is not limited to TMB solution, ABTS solution；Sample-loading buffer includes but is not limited to the Tris- containing bromophenol blue
HCl buffer solution；Enzyme labelled antibody is the antibody marked containing enzymes such as horseradish peroxidase, alkaline phosphatases, as HRP enzyme mark is anti-
Body；Standard items include but is not limited to cadmium chelating type CIC ELISA of the invention, other be chelated with the immune complex of cadmium；
Positive control include but is not limited to cadmium chelating type CIC ELISA of the present invention, other be chelated with the immune complex of cadmium, it is negative
Property control be dilution buffer.
Another object of the present invention is to provide a kind of method of quantitative detection cadmium chelating type CIC ELISA, with
Know that the above-mentioned cadmium chelating type immune complex of content as standard items, is detected using a pair of sample of following methods：It is enzyme-linked
Immunization, enzyme linked immunological and atomic absorption spectrum combined techniques, enzyme linked immunological and inductivity coupled plasma mass spectrometry combined techniques, purification
Immune complex and atomic absorption spectrum combined techniques, purification immune complex and inductivity coupled plasma mass spectrometry combined techniques, electricity
Swimming and enzyme linked immunological or atomic absorption spectrum or inductivity coupled plasma mass spectrometry combined techniques.
In the present invention, the having of can listing of method with detection cadmium chelating type immune complex is following several, but not
It is limited to following several.
Unless otherwise indicated, the laboratory operating procedures being related to are the step of this field routine to the present invention, and reagent, material are such as
It is following cited, do not enumerate in the present invention come be commonly used in the art or can be obtained by commercially available mode：
Dilution buffer is the 0.05M carbonate buffer solution of pH9.6, preparation method example：Take the Na of 1.5g2CO3With
2.93g NaHCO3Dissolution plus ddH2O is settled to 1000mL；
Washing buffer is the 0.15MPBS buffer of pH7.4, preparation method example：Take the KH of 0.2g2PO4, 2.90g
Na2HPO4·12H2O, KCl, 0.5mLTween-20 of NaCl, 0.2g of 8.0g, dissolution plus ddH2O is settled to 1000mL；
Confining liquid is bovine serum albumin solution, preparation method example：0.1g bovine serum albumin(BSA) is taken, washing buffer is added
Liquid dilution is settled to 100mL；
Terminate liquid is 2MH2SO4, preparation method example：Take the ddH of 178.3mL2O, to ddH2It is added dropwise along wall in O dense
H2SO4, it is stirring while adding, it is settled to 200mL；
The pH of substrate buffer solution is 5.0, Na2HPO4Molar concentration be 0.2M, the molar concentration of citric acid is 0.1M, often
The preparation method of the substrate buffer solution of 50mL is as follows：Take 1.42gNa2HPO4, 0.96g citric acid, ddH is then added2O to 50mL,
To obtain the final product；
Substrate is methyl biphenyl amine (TMB) solution, and methyl biphenyl amine (TMB) solution is by the group distribution according to following ratio
It makes：TMB：Substrate buffer solution：0.75%H2O2=0.5mL：10mL：32 μ L, wherein TMB is the methyl biphenyl amine second of 2g/L
Alcoholic solution；
The albumen that immune complex can be captured is the albumen that can be specifically bound with immune complex, including but not limited to
Such as C1Q, CIF albumen, anti-C_3 antibody；In following embodiment, can capture immune complex albumen it is specifically used be
C1qRecombinantProtein, article No. are " NOVUSH00000712-p01 "；
The substance or anti-Cd2+ antibody that metal Cd2+ can be captured are to buy from Ba Ao moral biotech company (BioWorld
Inc) the Cadmium resistance antibody of article No. BNA0008 or the model AP7015 of Ba Ao get Biotechnology Co., Ltd production；Wherein, originally
Substance, the anti-Cd2+ antibody of invention and cadmium specific binding all can be same substances；
The antibody that can be specifically bound with people's carrier protein is rabbit-anti people carrier protein antibodies, is commercially available.
Dilution multiple proportions is w/v below.
Method one：ELISA method detection cadmium chelating type immune complex, specific step is as follows：
1) albumen that can capture immune complex is coated on solid phase carrier：C1Q is diluted with dilution buffer
Albumen to 2600-20000 times, be added elisa plate micropore in, 4 DEG C overnight 16-18 hour or 37 DEG C water-bath 1-3 hours, store
Refrigerator；
2) it closes：Dilution buffer is removed, and is washed with washing buffer, after the completion of washing, adds confining liquid, 37
It DEG C places 1 hour, removes confining liquid, and washed with washing buffer, after the completion of washing, elisa plate is placed 1 small in 37 DEG C
When；
3) add sample to be tested, and incubate：It is sampled from the circulatory system, makees sample to be tested；With the cadmium chelating type of known content
Immune complex makees standard items；With dilution buffer dilute 10-40 times, add in micropore, 37 DEG C effect 1-2 hours；
4) substance of capture cadmium is added, and incubates：Sample to be tested is removed, and is washed with washing buffer, it is to be washed
After the completion of washing, the dilution of addition dilution buffer has the substance of affinity with cadmium or can form antigen antibody complex with cadmium reaction
Cadmium resistance antibody be diluted to 32000-250000 times, 37 DEG C effect 1-2 hours, make itself and the cadmium reaction on immune complex；
5) enzyme conjugates incubates：Cadmium resistance antibody is removed, and is washed with washing buffer, after the completion of washing, is added
With the diluted HRP enzyme labelled antibody of dilution buffer, 37 DEG C effect 1-2 hours, make itself and Cadmium resistance antibody response；
6) substrate incubates：Enzyme labelled antibody is removed, and is washed with washing buffer, after the completion of washing, substrate is added,
37 DEG C are protected from light effect 30 minutes；
7) reaction is terminated：Terminate liquid is added dropwise to each micropore；
8) taking wavelength is 405nm, and elisa plate is placed in microplate reader to the OD for reading sample to be tested group and standard items respectively
Value draws standard curve, acquires the content of sample to be tested.
In this method, when step 8) detects, microplate reader can also not be used, qualitative inspection is directly carried out by colour developing situation.
This method utilizes enzyme linked immunosorbent assay (ELISA) (ELISA) principle, can be compound by the nospecific immunity in serum
Object extracts, and weight cadmium is partially chelated on the immune complex extracted, and the cadmium on this partial immunity compound can be with
The specific antibody of the substance or the Cadmium resistance that can form antigen antibody complex with cadmium reaction that are had affinity with cadmium captures, it
(antibody nonrecognition coating egg can be captured by the antibody that the enzymes such as horseradish peroxidase, alkaline phosphatase mark again afterwards
It is white), the antibody in capture reads OD value under the action of color developing agent and terminate liquid under instrument, and exempts from without containing chelating cadmium
Epidemic disease compound will not then be captured by the specific antibody of Cadmium resistance, will not be with the enzymes such as horseradish peroxidase, alkaline phosphatase
The antibody of label is captured, and also without containing cadmium (negative control group result is feminine gender) in agents useful for same, thus when read
When OD value is the positive as the result is shown, i.e., the provable cadmium for detecting to chelate on CIC ELISA.
Method two：ELISA method+AAS method detects cadmium chelating type immune complex, and specific step is as follows：
1) albumen that can capture immune complex is coated on solid phase carrier：Coating protein is diluted with dilution buffer
To 5800-15000 times, be added elisa plate micropore in, 4 DEG C overnight 16-18 hour or 37 DEG C water-bath 1-3 hours, storage refrigerator；
2) it closes：Dilution buffer is removed, and is washed with washing buffer, after the completion of washing, adds confining liquid, 37
It DEG C places 1 hour, removes confining liquid, and washed with washing buffer, after the completion of washing, elisa plate is placed 1 small in 37 DEG C
When；
3) add sample to be tested, and incubate：It is sampled from the circulatory system, makees sample to be tested；With the cadmium chelating type of known content
Immune complex makees standard items；With dilution buffer dilute 10-40 times, add in micropore, 37 DEG C effect 1-2 hours；
4) it elutes：Sample to be tested is removed, and is washed with washing buffer, after the completion of washing, eluent is added, in
It is acted on 1-2 hours at 37 DEG C；
5) it detects：It samples from ELISA micropore, chelates in Atomic Absorption Spectrometer detection in the cadmium on immune complex,
Read respective value.
This method further combines atomic absorption spectrum (AAS) principle on the basis of enzyme linked immunological principle, utilizes original
Sub- absorption spectrometer detection chelating is in the cadmium on CIC ELISA, due to only containing immune complex in solution, and it is used
Without cadmium (negative control group result be feminine gender) in reagent, result will not be shone into interference, thus when it is read as the result is shown
When for the positive, i.e., the provable cadmium for detecting to chelate on CIC ELISA.
The method of the present invention scope of application is wider, can be with save the cost, and increases the differential water pressures inside and outside bag filter, improves
It dialyses rate, can shorten manufacturing cycle, also the wastes such as ion in waste liquid are recycled, there is energy-saving and environment-friendly spy
Point avoids chemical contamination, therefore the present invention has the biggish market competitiveness.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (9)
1. a kind of excellent effect cadmium chelating type immune complex, which is characterized in that the cadmium chelating type immune complex is with next
Kind:
Cadmium ion is incorporated into the compound of immune complex formation；
Or cadmium ion be incorporated into after carrier protein with and the antibody that specifically binds of the carrier protein be formed by compound；
Or cadmium ion is incorporated into the compound formed in conjunction with carrier protein after immunoglobulin.
2. cadmium chelating type immune complex according to claim 1, it is characterised in that:The immune complex or carrier egg
At least contain one of zinc fingers, sulfydryl, cysteine residues, cadmium ion and immune complex or carrier in white structure
Albumen is combined with zinc fingers or sulfydryl or cysteine residues.
3. cadmium chelating type immune complex according to claim 2, it is characterised in that:The carrier protein is antibody egg
Among white, lipoprotein, hemoglobin, nuclear antigen, foreign sera antigen, virus, bacterium, protozoon, worm or immunoglobulin
It is a kind of.
4. cadmium chelating type immune complex according to claim 1, it is characterised in that:The cadmium ion is by chelating agent and carries
After body protein combination again in conjunction with energy and the antibody of carrier protein specific binding, the chelating agent is ITCBE, EDTA, more phosphorus
One of hydrochlorate, amino carboxylic acid, 1,3- diketone, hydroxycarboxylic acid.
5. a kind of preparation method of cadmium chelating type immune complex as described in claim 1, which is characterized in that including following step
Suddenly：
S1：First chelating agent solution is prepared and is completed, then chelating agent is dissolved in solvent, chelating agent solution can be obtained；
S2：Appropriate carrier protein is taken, carrier protein is added in borate buffer solution, can be prepared by carrier protein solution；
S3：Chelating agent solution obtained is added in carrier protein solution, is placed in water bath with thermostatic control and stirs evenly, constant temperature
Water-bath 4-30h, can be obtained mixed liquor；
S4：It fetches bag filter and EDTA-NaHCO is added in bag filter3Solution boils, and bag filter is fixed on to the middle part of water tank,
And enough ddH are added in water tank2O, the ddH2The volume ratio of solution is 85-90 in the volume and bag filter of O:1, repeating should
Mixing in S3 is fitted into bag filter by step 2-4 times, and bag filter is placed in constant temperature water tank, and foot is added in constant temperature water tank
The ddH of amount2O, the ddH2The volume ratio of solution is 85-90 in the volume and bag filter of O:1, it, will be in bag filter after dialysed overnight
Liquid collect.
S5：PH to 7.0 is adjusted dilute hydrochloric acid solution is added from the liquid collected in bag filter, is slowly dropped into cadmium-ion solution,
Do not stop to shake when instillation, instills and complete to be placed in water bath with thermostatic control and stir evenly, water bath with thermostatic control 4-30h, then repeatedly S4 mono-
It is secondary, liquid is collected, cadmium chelating type antigen is obtained.
S6：The waste liquid collected in solution and S5 in S4 outside bag filter is mixed, is added and changes into mixed solution
Property the cellulosic material crossed adsorbed, and repeat to add the above-mentioned cellulosic material being modified, reuse atomic absorption method pair
Processed solution carries out quantitative analysis, and when cadmium ion enrichment degree reaches 0.1mol/ml in solution, recycling cadmium ion is sharp again
With.
S7：The antibody that can be specifically bound with carrier protein is added in above-mentioned cadmium chelating type antigen, cadmium chelating is obtained after reaction
Type immune complex.
6. cadmium chelating type immune complex as described in claim 1 is in the reagent of preparation detection cadmium chelating CIC ELISA
Or the application in enzyme linked immunological kit.
7. a kind of enzyme linked immunological kit of detection cadmium chelating CIC ELISA, it is characterised in that：The kit includes such as
The standard items of cadmium chelating type immune complex described in claim 1.
8. enzyme linked immunological kit as claimed in claim 6, which is characterized in that the kit further includes that at least one or less is tried
Agent：The coating buffer of albumen containing capture antigen antibody complex or the coating buffer of the antibody containing capture cadmium, enzyme labelled antibody.
9. a kind of method of qualitative detection cadmium chelating type CIC ELISA, which is characterized in that wanted with the right of known content
Cadmium chelating type immune complex described in asking 1 is detected as standard items using a pair of sample of following methods：Enzyme linked immunological
Method, enzyme linked immunological and atomic absorption spectrum combined techniques, enzyme linked immunological and inductivity coupled plasma mass spectrometry combined techniques, purification are immune
Compound and atomic absorption spectrum combined techniques, purification immune complex and inductivity coupled plasma mass spectrometry combined techniques, electrophoresis and
Enzyme linked immunological or atomic absorption spectrum or inductivity coupled plasma mass spectrometry combined techniques.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810594250.0A CN108828207A (en) | 2018-06-11 | 2018-06-11 | A kind of excellent effect cadmium chelating type immune complex and preparation method thereof |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US5639624A (en) * | 1989-03-14 | 1997-06-17 | Board Of Regents Of The University Of Nebraska | Monoclonal antibodies specific for metallic cations and method therefor |
| JPH11174054A (en) * | 1997-12-08 | 1999-07-02 | Tadayuki Imanaka | Measuring method for heavy metal and recovery method therefor |
| CN102725008A (en) * | 2009-08-07 | 2012-10-10 | 汉莫堤克股份有限公司 | Device and method for eliminating biologically harmful substances from bodily fluids |
| CN104965076A (en) * | 2015-07-14 | 2015-10-07 | 上海拜豪生物科技有限公司 | Cadmium-chelated immune complex as well as preparation method and application thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5639624A (en) * | 1989-03-14 | 1997-06-17 | Board Of Regents Of The University Of Nebraska | Monoclonal antibodies specific for metallic cations and method therefor |
| JPH11174054A (en) * | 1997-12-08 | 1999-07-02 | Tadayuki Imanaka | Measuring method for heavy metal and recovery method therefor |
| CN102725008A (en) * | 2009-08-07 | 2012-10-10 | 汉莫堤克股份有限公司 | Device and method for eliminating biologically harmful substances from bodily fluids |
| CN104965076A (en) * | 2015-07-14 | 2015-10-07 | 上海拜豪生物科技有限公司 | Cadmium-chelated immune complex as well as preparation method and application thereof |
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