CN103627677A - Lp(a) (Lipoprotein(a)) resisting monoclonal antibody and latex-enhanced immunonephelometric detection kit for Lp(a) - Google Patents
Lp(a) (Lipoprotein(a)) resisting monoclonal antibody and latex-enhanced immunonephelometric detection kit for Lp(a) Download PDFInfo
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Abstract
The invention discloses an Lp(a) (Lipoprotein(a)) resisting monoclonal antibody and a latex-enhanced immunonephelometric detection kit for Lp(a). The invention provides a hybridoma cell strain which can stably secrete the Lp(a) monoclonal antibody and has the collection number of CGMCC No. 8509. The hybridoma cell strain provided by the invention can stably produce the Lp(a) monoclonal antibody, and personal pairing, specific Lp(a) combination and no combination with Kringle IV-2 can be realized. Shown by a test that serum samples of patients are detected by an Lp(a) kit prepared from the Lp(a) resisting monoclonal antibody, values detected by the latex-enhanced immunonephelometric detection kit prepared from the Lp(a) resisting monoclonal antibody have high conformity with detected values of commercially available kits, and the accuracy is reliable, so that the latex-enhanced immunonephelometric detection kit can be applied to Lp(a) detection.
Description
Technical field
The present invention relates to the monoclonal antibody of hybridoma cell strain and secretion, relate in particular to a kind of anti-Lp(a) [Lipoprtein (a), Lp (a)] monoclonal antibody and the hybridoma cell strain of secreting this monoclonal antibody, the further Lp of the present invention (a) latex enhancing immune, than turbid detection kit, belongs to the detection field of lipoprotein (a).
Background technology
Lipoprotein (a) ［ Lipoprotein (a), Lp (a) ］ be the lipoprotein of a kind of uniqueness in human body, by low-density lipoprotein (LDL) sample particulate and lipophorin (a) ［ apolipoprotein (a), apo (a) ］, formed.The gene height homology of Apo (a) and Profibrinolysin, belongs to human plasmin protogene superfamily member.Apo (a) consists of the Repeat of the plasminogen Kringle IV sample of multiple copied and Kringle V and the serine protease region of single copy.Apo (a) has the motif (called after Kringle IV-1～Kringle IV-10) of 10 kinds of plasminogen Kringle IV samples, and except Kringle IV-2, every kind of Kringle IV is single copy.Apo (a) polymorphism is subject to Gene Handling, the copy number of different apo (a) hypotype Kringle IV-2 be 3-40 not etc.About the maximum epidemiological study of Lp (a), proved that Lp (a) level and heart disease and cerebral infarction exist continuous independently moderate relevant, high Lp (a) gains public acceptance as the independent hazard factor of cardiovascular and cerebrovascular atherosclerosis.
Lp (a) concentration detects conventional method double antibody sandwich ELISA, immunoturbidimetry and latex enhancing immune turbidimetry.These three kinds of methods all need to use Lp (a) antibody, wherein latter two method in Clinical Laboratory, apply more, latex enhancing immune turbidimetry particularly.Because Lp (a) molecular weight is large, complex structure, also cannot prepare Lp (a) antigen by artificial expression means at present.Polyclonal antibody (how anti-) preparation with Lp (a) antigen can only be from serum purification, because Lp (a) has polymorphism between different people, be subject to the restriction in serum source, the more difficult control of difference between batch of Lp (a) antigen, finally causes the how anti-difference between batch of Lp (a) larger.In addition, due to the difference of apo (a) molecule Kringle IV-2 copy number, the impact of using monoclonal antibody or the how anti-test kit of preparing as raw material for this site can be subject to antigenic heterogeneity, causes the inaccurate of detected result.
Polyclonal antibody (how anti-) preparation with Lp (a) antigen can only be from serum purification, because Lp (a) has polymorphism between different people, be subject to the restriction in serum source, the more difficult control of difference between batch of Lp (a) antigen, finally causes the how anti-difference between batch of Lp (a) larger.Monoclonal antibody (monoclonal antibody) is not although rely on Lp (a) antigen, existing monoclonal antibody or identification pleomorphism site, or need to match use with other monoclonal antibodies.The latex enhancing immune of take is example than turbid reagent: in reagent preparation, the corresponding coupling operation of increase meeting of antibody type and concentration are adjusted operation, and workload is doubled and redoubled, and the risk that the mutation of test kit poor performance is large also can produce thus.
The monoclonal antibody of epitope beyond exploitation identification apo (a) molecule Kringle IV-2, can fine solution antibody difference between batch the large and polymorphic sex problem of apo (a).
Summary of the invention
One of object of the present invention is to provide a strain can produce self pairing, specific binding Lp (a), and the hybridoma cell strain of energy stably excreting antibody;
Two of object of the present invention is to provide the monoclonal antibody of being secreted by described hybridoma cell strain;
A further object of the present invention is this monoclonal antibody to be applied to detect Lp (a) and to provide a kind of Lp (a) latex enhancing immune than turbid detection kit.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
The inventor is screening antigen with the polymorphic bands Kringle IV-2 in Lp (a) molecule, adopt indirect elisa method to carry out the screening of Lp (a) monoclonal antibody specific, by chessboard method, carry out the pairing screening of monoclonal antibody, finally obtained 1 strain can produce self pairing, specific binding Lp (a) and with the monoclonal antibody target cell strain of Kringle IV-2 without combination, thereby completed the present invention.
First the present invention provides a strain to produce can self pairing, specific binding Lp (a) and with the hybridoma cell strain of Kringle IV-2 without anti-Lp (a) monoclonal antibody of combination.
The present invention is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center by the screen hybridoma cell strain obtaining, deposit number: CGMCC No.8509; Classification And Nomenclature is: the strain of Lp(a) monoclonal antibody hybridoma cell; Preservation date: on November 20th, 2013; Depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
The present invention also provides a kind of method of preparing described hybridoma cell strain, comprise: with the polymorphic bands Kringle IV-2 in Lp (a) molecule, be screening antigen, adopt indirect elisa method and double antibodies sandwich ELISA method to carry out the operation of Lp (a) monoclonal antibody specific screening, screening obtains described hybridoma cell strain.
Hybridoma cell strain in the present invention and anti-Lp (a) monoclonal antibody can produce by method hereinafter described.Further, those skilled in the art can use these according to below describing and the transformed method being more suitable for of techniques known in the art.
(1) immunogenic preparation
It is immunogen that the present invention adopts Lp (a) molecule of natural extract purifying.Can be that Lp (a) calibration object antigen is through molecular sieve, the separation and purification of the S-200 of RuGE company filler obtains, also can be commercial high purity Lp (a) natural antigen, can also from the higher serum of Lp (a) content, by literature method, be prepared.
(2) preparation of detectable antigens
Detectable antigens comprises Lp (a) natural antigen, APOB antigen and Lp (a) Kringle IV-2 prokaryotic expression antigen.Lp (a) natural antigen, APOB antigen can obtain by purchase or other modes.Lp (a) Kringle IV-2 obtains by Prokaryotic expression, purification, concrete is: the aminoacid sequence (SEQ ID No.2) that obtains Kringle IV-2 from NCBI website, by the anti-codon gene order (SEQ ID No.1) that is translated into intestinal bacteria preference of Vector NTI software, sending outsourcing company to carry out full gene synthesizes, insert in expression vector, transform e. coli bl21 (DE3), carry out abduction delivering and protein purification.
(3) preparation of monoclonal antibody
Lp (a) molecular weight is large, and easily induction produces antibody, and the present invention adopts Lp (a) natural antigen immune mouse, carries out 3-5 immunity.Collect splenocyte after immunization 2-5 days the last time (preferably 3 days), the cell that produces antibody is merged to rear selection with myeloma cell and Lp (a) has the hybridoma of specific binding in these tissues, thereby prepare the cell of product monoclonal antibody.
Fusion can be carried out according to known method, for example can enumerate following method: splenocyte is mixed in the ratio of 5-10:1 with oncocyte, centrifugal going after supernatant adds fusion to promote reagent in throw out, then with DMEM substratum, stops merging the effect of promotion reagent.The centrifugal cell of abandoning after supernatant is resuspended with the HAT substratum of suitable volumes, makes fused cell, and fused cell is cultivated at the Kong Gezhong that cultivates hybridoma.For myeloma cell, can use known myeloma cell, clone such as enumerating the mouse sources such as SP2/0-Ag14 (SP2/0), P3/X63-Ag8 (X63), P3/X63-Ag8.653 (X63-Ag8.653), P3/NSI-1-Ag4-1 (NS-1), is wherein preferably SP2/0.Merge and promote that reagent can be polyoxyethylene glycol
1450(PEG
1450), polyoxyethylene glycol
4000(PEG
4000) etc., be preferably PEG
1450.
The selection of monoclonal antibody can be carried out according to known method.Usually to carry out at the cell culture medium for zooblast that is added with HAT.Substratum for selection and culture for example can be enumerated: volume ratio is DMEM or the RPMI-1640 substratum of 5%-20% foetal calf serum.Cell generally in the environment of 5-10%CO2 gas in 36.5 ℃-37.5 ℃, cultivate 10-14 days.
From cultivating the Kong Gezhong of hybridoma, collect the supernatant liquor of culture, the culture hole of secretion object antibody can be selected by indirect ELISA method.First, 3 kinds of antigens such as the reaction of Lp (a) natural antigen, APOB antigen and Kringle IV-2 are fixed in 96 hole enzyme plates, and with tween 20 phosphate buffered saline buffer (PBST) detersive enzyme target, then culture supernatant sample is joined in different antigen coated holes, 37 ℃ of incubations are after 30 minutes, sheep anti mouse-the HRP as ELIAS secondary antibody that adds 1:10000 dilution, 37 ℃ of incubations add Urea Peroxide-tetramethyl benzidine substrate (H after 30 minutes
2o
2-TMB) as substrate, develop the color, with after sour termination reaction, measure the light absorption value under 450nm.
Choose with Lp (a) natural antigen and react, and with APOB antigen and the unresponsive culture hole of Kringle IV-2, by known limiting dilution assay, clone.By cultivating the anti-Lp of acquisition (a) monoclonal antibody that this hybridoma cell strain can be a large amount of.The acquisition methods of this anti-Lp (a) monoclonal antibody, the earth can be divided into two kinds.Be to use a substratum, in the culture vessels such as flask, cultivate, from this supernatant liquor, obtain the method for antibody.It for volume ratio, is for example the DMEM nutrient solution of 10%-20% foetal calf serum.Another kind method be by the hybridoma cell strain of cultivating with culture vessel by the animal that induces method in Syngenic mice body and be inoculated into homology, obtain the monoclonal antibody of anti-Lp (a).
Employing indirect elisa method of the present invention carries out in the operation of screening of Lp (a) monoclonal antibody specific,
Comprise antigen antigen is fixed in 96 hole enzyme plates, and with the coated elisa plate of tween 20 phosphate buffered saline buffer (PBST) detersive enzyme target; Culture supernatant sample is joined in different antigen coated holes, and 37 ℃ of incubations, after 30 minutes, are used the operation of the interpolation culture supernatant sample of PBST detersive enzyme target; Sheep anti mouse-the HRP as ELIAS secondary antibody that adds 1:10000 dilution, 37 ℃ of incubations are used the operation of the interpolation ELIAS secondary antibody of PBST detersive enzyme target after 30 minutes; And the operation developing the color.
Another object of the present invention is to provide a kind of anti-Lp (a) monoclonal antibody being produced by this hybridoma cell strain.Hybridoma cell strain of the present invention can stably be produced anti-Lp (a) monoclonal antibody, and anti-Lp (a) monoclonal antibody being produced by this hybridoma cell strain can realize self pairing, specific binding Lp (a), and with Kringle IV-2 without combination.Because this anti-Lp (a) monoclonal antibody can meet the requirement that Lp (a) detects well, therefore can be applied in Lp (a) detection kit.
Therefore, the invention provides a kind of Lp (a) latex enhancing immune than turbid detection kit, by reagent 1(R1 independent of each other) and reagent 2(R2) form; Wherein, the composition of R1 reagent 1 comprises damping fluid, sanitas etc.; The composition of reagent 2 comprises that damping fluid, bovine serum albumin (BSA) or calf serum (NCS), coupling have the latex microsphere of anti-Lp (a) monoclonal antibody etc.
Concrete, the composition of reagent 1 can be: 1) 10-50mM, pH7.4 phosphate buffered saline buffer (PBS), 0.02%-0.1% sodium azide (NaN3); Or 2) 50-200mM, pH7.4Tris-HCl damping fluid, 0.02%-0.1%NaN3; Or 3) 50-200mM, pH7.4MOPS damping fluid, 0.02%-0.1%NaN3.Be preferably: 20mM, pH7.4 phosphate buffered saline buffer, 0.1%NaN3.
The composition of reagent 2 can be, 1) and 10-50mM, pH7.4 phosphate buffered saline buffer, 0.1-0.5%BSA or 5-15%NCS, 0.125%-0.25% coupling has latex microsphere (the antibody coupling ratio: 0.5-2mg/10mg latex microsphere of antibody; Latex microsphere particle diameter: 80-200nm); Or 2) 50-200mM, pH7.4Tris-HCl damping fluid, 0.1-0.5%BSA or 5-15%NCS, 0.125%-0.25% coupling has latex microsphere (the antibody coupling ratio: 0.5-2mg/10mg latex microsphere of antibody; Latex microsphere particle diameter: 80-200nm); Or 3) 50-200mM, pH7.4MOPS damping fluid, 0.1-0.5%BSA or 5-15%NCS, 0.125%-0.25% coupling has latex microsphere (the antibody coupling ratio: 0.5-2mg/10mg latex microsphere of antibody; Latex microsphere particle diameter: 80-200nm); Be preferably: 20mM, pH7.4 phosphate buffered saline buffer, 0.1%BSA, 0.2% coupling has latex microsphere (the antibody coupling ratio: 1mg/10mg latex microsphere of antibody; Latex microsphere particle diameter: 124nm).
Hybridoma cell strain of the present invention can stably be produced anti-Lp (a) monoclonal antibody, and anti-Lp (a) monoclonal antibody being produced by this hybridoma cell strain can realize self pairing, specific binding Lp (a), and with Kringle IV-2 without combination.The test that LPa-3 Lp that monoclonal antibody is joined (a) test kit detects patients serum's sample shows, detection kit and the Lp of company of DESAY (a) test kit measured value degree of conformity by the preparation of anti-Lp of the present invention (a) monoclonal antibody are high, accuracy is reliable, can be applied in Lp (a) test kit.
Term definition involved in the present invention
The present invention said " Lp (a) " unless stated otherwise, refers to Lp(a) and associated products thereof in human serum or blood plasma, as lipoprotein (a) precursor, mature form or fragment.
The present invention's said " specific binding " refer to anti-Lp (a) monoclonal antibody only with Lp (a) combination, and not with apolipoprotein B (APOB) combination.
The present invention's said " antibody " refer to can with Lp (a) molecule of complete overall length or the monoclonal antibody that Lp (a) fragments specific is combined, or the Partial Fragment of these antibody.
Said self pairing of the present invention refers to that a kind of monoclonal antibody can be attached to two or more forms the phenomenon on same antigen molecule, and the reason that produces this phenomenon is on antigen molecule, to have two or more same or analogous epitopes.
Accompanying drawing explanation
Fig. 1 is that the monoclonal antibody being produced by LPa-3 hybridoma cell strain is applied to the correlation analysis figure in test kit.
Specific embodiment
Below in conjunction with specific embodiments, further describe the present invention, advantage and disadvantage of the present invention will be more clear along with description.But these embodiment are only exemplary, scope of the present invention are not formed to any restriction.It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can the details of technical solution of the present invention and form be modified or be replaced, but these modifications and replacement all fall within the scope of protection of the present invention.
The structure of embodiment 1 hybridoma cell strain
With 4 of Lp (a) antigen immunes female Balb/c mouse in 6 week age, first immunisation adopts Freund's complete adjuvant that antigen is carried out to emulsification, every injected in mice 40ug antigen, then adopts Freund's incomplete adjuvant emulsification antigen, take and within 21 days, as interval, carries out the 2-3 time immunity.Complete 3 immune posterior orbit blood samplings and detect serum antibody titer, choose the higher mouse of tiring and carry out cytogamy.Merge and with Lp (a) antigen that does not add adjuvant, selected mouse is impacted to immunity in first 3 days, after 3 days, the disconnected cervical vertebra of mouse is put to death, aseptic separating spleen, prepare splenocyte suspension, the SP2/0 cell of splenocyte and vitro culture is mixed in 10:1 ratio, after centrifuge washing, with PEG1450, carry out cytogamy, fused cell is cultivated 8 days, adopt indirect elisa method to screen fused cell culture supernatant, detectable antigens is Lp (a), APOB and Kringle IV-2.With 0.05M pH9.6Na2CO3-NaHCO3 damping fluid, Lp (a), APOB and Kringle IV-2 are diluted to 1ug/ml, three kinds of antigens intersect coated by row, every hole 100ul is added in enzyme plate, 4 ℃ of placements are spent the night, with 0.15M pH7.4, contain 0.05% tween 20 phosphate buffered saline buffer (PBST) detersive enzyme target 1 time, complete antigen coated; Culture supernatant is added in enzyme plate, and each culture hole sampling 3 times, is added to respectively the coated hole Zhong，Mei hole 60ul of Lp (a), APOB and KringleIV-2, and 37 ℃ of incubation 30min, wash plate 2 times with PBST; HRP-goat anti-mouse igg is diluted to working concentration, and every hole 100ul is added in enzyme plate, and 37 ℃ of incubation 30min wash plate 3 times with PBST; Add TMB-H2O2 substrate, 37 ℃ of lucifuge colour developing 10min; Every hole adds 50ul2M H2SO4 color development stopping, reads the light absorption value of 450nm in microplate reader.Choose with Lp (a) and be strong positive reaction, but the culture hole cloning that is negative and reacts with APOB and Kringle IV-2, and set up hybridoma cell strain.
The pairing screening of the anti-Lp of embodiment 2 (a) monoclonal antibody
Clone is built to Lp (a) the monoclonal antibody hybridoma of strain and prepare Lp (a) odd contradictive hydroperitoneum by ascites method, with Protein G column purification, go out Lp (a) monoclonal antibody, sodium periodate method is carried out HRP mark to Lp (a) monoclonal antibody, then by chessboard method, the pairing characteristic of each strain Lp (a) monoclonal antibody is analyzed.Each strain Lp (a) monoclonal antibody is diluted to 5ug/ml with 0.05M pH9.6Na2CO3-NaHCO3 damping fluid, every hole 100ul coated elisa plate, every kind of coated 1 row hole bar of monoclonal antibody; With PBST, Lp (a) antigen diluent is become to 0.1ug/ml, every hole 100ul, 37 ℃ of incubation 30min, wash plate 2 times with PBST; With PBST, the HRP marker of each strain monoclonal antibody is diluted to working concentration, every kind of monoclonal antibody marker adds 1 row's plate hole, every hole 100ul, and 37 ℃ of incubation 30min, wash plate 3 times with PBST; Add TMB-H2O2 substrate, 37 ℃ of lucifuge colour developing 10min, every hole adds 50ul2M H2SO4 color development stopping, reads OD450 value in microplate reader.The coated monoclonal antibody that positive reaction hole is corresponding and HRP mark monoclonal antibody are the monoclonal antibody of pairing.Choose the good monoclonal antibody of pairing effect and make the Preliminary Applications of sandwich ELISA method.The data of monoclonal antibody pairing screening are in Table 1
Table 1Lp (a) monoclonal antibody chessboard method pair analysis result
Lp (a) detection kit of embodiment 3 use LPa-3 monoclonal antibody preparations
1) reagent 1 is:
PBS damping fluid (pH7.4)
BSA 0.5%
Tween 20 0.1%
NaN3 0.1%
2) reagent 2 is:
PBS damping fluid (pH7.4)
3) preparation of LPa-3 monoclonal antibody latex microsphere
A) with 50mM HEPES damping fluid (pH7.4) by LPa-3 monoclonal antibody to 1mg/ml.
B) with MES damping fluid (10mM pH5.0), diluting 100nm latex microsphere to mass concentration is 2%, totally 400 μ l.
C) with MES damping fluid (10mM pH5.0), prepare respectively EDC(2mg/ml) 400 μ l, NHS (2mg/ml) 400 μ l.
D) in latex microsphere solution, dropwise add 300 μ l EDC solution, then dropwise add 300 μ l NHS solution, stirring at room 15min.
E) microballoon of activation and monoclonal antibody are mixed to stirring at room 2 hours.
F) centrifugal after reaction, remove supernatant, precipitation is resuspended with 6ml reagent 2 damping fluids, ultrasonic dispersion.
Embodiment 4LPa-3 Lp that monoclonal antibody is joined (a) test kit detects the test of patients serum's sample
With LPa-3 Lp that monoclonal antibody is joined (a) test kit, patients serum's sample of 30 parts of Different L p (a) content is measured on automatic clinical chemistry analyzer Olympus AU400, with the Shanghai Lp of company of Shen Neng DESAY (a) latex enhancing immune than turbid test kit in contrast, measured value is the content of Lp (a), its unit is mg/dL, and the content of normal human serum Lp (a) is lower than 30mg/dL.Two kinds of measurement results are contrasted, the results are shown in table 2.Wherein, Self-made reagent box survey location parameter is: R1:R2:S=180ul:60ul:2ul, contrast agents box by specification parameter is set.Meanwhile, Self-made reagent He Yu DESAY commercially available reagent box detected result is done to correlation analysis curve, its result is referring to Fig. 1.
From table 2 and Fig. 1, the detected result of Self-made reagent box and the Lp of company of DESAY (a) test kit good relationship: dependency equation is y=1.08x-6.3519, R
2=0.9933.From this result, high by test kit and the Lp of company of DESAY (a) the test kit measured value degree of conformity of the preparation of anti-Lp of the present invention (a) monoclonal antibody, accuracy is reliable, can be applied in Lp (a) test kit.
Table 2LPa-3 monoclonal antibody Self-made reagent He Yu DESAY test kit comparison data
Claims (8)
1. the hybridoma cell strain of the anti-Lp of a strain stably excreting (a) monoclonal antibody, is characterized in that, its microbial preservation number is: CGMCC No.8509.
2. a method that builds hybridoma cell strain described in claim 1, it is characterized in that, comprise: with the polymorphic bands Kringle IV-2 in Lp (a) molecule, be screening antigen, adopt indirect elisa method and double antibodies sandwich ELISA method to carry out the screening of Lp (a) monoclonal antibody specific and obtain described hybridoma cell strain.
3. anti-Lp (a) monoclonal antibody of being secreted by hybridoma cell strain claimed in claim 1.
4. anti-Lp claimed in claim 3 (a) monoclonal antibody detects the purposes in lipoprotein (a) reagent in preparation.
5. detect a test kit for lipoprotein (a), it is characterized in that: contain anti-Lp claimed in claim 3 (a) monoclonal antibody.
6. Lp (a) latex enhancing immune, than a turbid detection kit, is comprised of with reagent 2 reagent 1 independent of each other; Wherein, the composition of reagent 1 comprises damping fluid, sanitas; The composition of reagent 2 comprises that damping fluid, bovine serum albumin or calf serum, coupling have the latex microsphere of anti-Lp (a) monoclonal antibody; It is characterized in that: described anti-Lp (a) monoclonal antibody is that the hybridoma cell strain of claim 1 is secreted.
7. according to Lp claimed in claim 6 (a) latex enhancing immune, than turbid detection kit, it is characterized in that: the composition of reagent 1 comprises: 20mM, pH7.4 phosphate buffered saline buffer, 0.1%NaN3.
8. according to Lp claimed in claim 6 (a) latex enhancing immune, than turbid detection kit, it is characterized in that: the composition of reagent 2 is: 20mM, pH7.4 phosphate buffered saline buffer, 0.1%BSA, 0.2% coupling has the latex microsphere of antibody; Antibody coupling ratio wherein: 1mg/10mg latex microsphere; Latex microsphere particle diameter is 124nm.
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| CN104597252A (en) * | 2015-01-23 | 2015-05-06 | 浙江卓运生物科技有限公司 | Kit for detecting oxidized low-density human serum lipoproteins by immunological turbidimetry |
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| CN106501536A (en) * | 2016-10-03 | 2017-03-15 | 王贤俊 | A kind of latex orientation coupling technology detection lipoprotein（a）Test kit |
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| CN106990234B (en) * | 2017-05-31 | 2019-03-26 | 吉林省汇酉生物技术股份有限公司 | A kind of detection reagent and method of lipoprotein (a) |
| CN111239422A (en) * | 2018-12-25 | 2020-06-05 | 武汉生之源生物科技股份有限公司 | Serum lipoprotein (a) latex enhanced turbidimetric detection kit |
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