CN103525769B - Hybridoma and the application of anti-retinol conjugated protein monoclonal antibody can be secreted - Google Patents
Hybridoma and the application of anti-retinol conjugated protein monoclonal antibody can be secreted Download PDFInfo
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Abstract
The present invention relates to a kind of hybridoma secreting anti-retinol conjugated protein monoclonal antibody, does is its preserving number CCTCC? No:C2013132.The secretion output of this hybridoma is high, it is secreted the anti-retinol conjugated protein monoclonal antibody obtained and has the advantage such as high-affinity, high specific, Retinal-binding protein detection field can be widely used in, as the preparation field etc. of detection reagent or test set, in specificity, sensitivity and verification and measurement ratio etc., than traditional detection method, there is significant advantage.In addition, the invention still further relates to the application of the monoclonal antibody of this hybridoma a kind of secretion, this hybridoma and monoclonal antibody thereof.
Description
Technical field
The present invention relates to retinol conjugated protein field of immunodetection, especially relate to a kind of hybridoma and application of secreting anti-retinol conjugated protein monoclonal antibody.
Background technology
β2-microglobulin and RBP(retinol conjugated protein is had clinically at present) for the urine small molecular protein of renal dysfunction early diagnosis.β2-microglobulin is very unstable when pH ﹤ 5.5, easily degrades, and the transformation period of β2-microglobulin in urine is shorter, and in pH4.0 environment, placement has the degraded of 70% after 4 hours, and RBP only has degraded 5% under the same conditions.Therefore, RBP is more stable in renal dysfunction diagnosis relative to β2-microglobulin, more reliably.
RBP is a kind of low molecular protein, and molecular weight is about 21KD, and primarily of liver cell synthesis, major function also can be combined with retinal epithelium tissue specificity, for it provides vitamin A from hepatic transport to epithelial cell by Vogan-Neu.The RBP-Vogan-Neu polymkeric substance secreted in liver and Transferrins,iron complexes form stable albumen composition and circulate in blood system.RBP-Vogan-Neu-Transferrins,iron complexes complex body disintegrates after Vogan-Neu release, and RBP is present in blood system in a free form, by glomerular filtration, is more heavily absorbed by proximal tubular epithelial cells, degrades.
RBP free in blood can from glomerular filtration, wherein 99.9% heavily to absorb through proximal tubular epithelial cells, degrades.Under normal circumstances, RBP excretion very micro-(lower than 300ng/mL) in urine.If uriniferous tubules pathology, then heavy absorptive function declines, and the RBP in urine can be caused to rise in a large number.In injury of renal tubular patient urine, RBP content is greater than 300ng/mL, generally can reach more than 5 times of normal people, and the RBP content in its urine of severe patient even can reach 10000ng/mL.Therefore in urine, RBP increases and has early diagnosis meaning, is considered to a good early sign thing of injury of renal tubular, has been widely used in clinically in the diagnosis of kidney injury the detection of RBP.
The clinical detection meaning of RBP also comprises acute glomerulonephritis, chronic glomerulonephritis, diabetic nephropathy and chronic renal failure disease etc., and the concentration level of its serum RBP all raises.Simultaneously RBP and hepatopathy also have relation, all remarkable negative correlation of RBP and serum bilirubin, glutamic-oxal(o)acetic transaminase and alkaline phosphatase activity, and the serum RBP level of gangrenosum acne liver cirrhosis, cholehepatocirrhosis and acute hepatitis, chronic hepatitis patient is all remarkable in Normal group.
The measuring method of traditional RBP mainly contains euzymelinked immunosorbent assay (ELISA), radioimmunology and latex immunoturbidimetry.Immunoturbidimetry, because of its stable reagent, easily realizes high-throughput and the operation automation of detection, and detected result is quick, accurate, reliable, therefore becomes the measuring method of clinical labororatory's most prospect.
Summary of the invention
Based on this, be necessary to provide a kind of hybridoma secreting anti-retinol conjugated protein monoclonal antibody.
In addition, there is a need to the application providing the monoclonal antibody of above-mentioned hybridoma or its secretion in medical science.
Secrete a hybridoma for anti-retinol conjugated protein monoclonal antibody, preserving number is CCTCCNo:C2013132.
The above-mentioned hybridoma secreting anti-retinol conjugated protein monoclonal antibody is in preparation diagnosis kidney injury disease or the detection reagent of hepatic diseases or the application in test set field.
The above-mentioned hybridoma secreting anti-retinol conjugated protein monoclonal antibody is in the application in preparation Retinal-binding protein detection reagent or test set field.
A kind of monoclonal antibody of being secreted by the above-mentioned hybridoma secreting anti-retinol conjugated protein monoclonal antibody.
Said monoclonal antibody is in preparation diagnosis kidney injury disease or the detection reagent of hepatic diseases or the application in test set field.
Said monoclonal antibody is preparing the application of Retinal-binding protein detection reagent or test set field.
A kind of Retinal-binding protein detection test kit, comprises said monoclonal antibody.
Wherein in an embodiment, described Retinal-binding protein detection test kit comprises the first reagent and the second reagent, wherein, described first reagent comprises 30-60mmol/LTris damping fluid, 60-95mmol/L polyoxyethylene glycol, 1-5mmol/L sodium azide, 6-13mmol/L sodium ethylene diamine tetracetate and 1-5mmol/L bovine serum albumin; Described second reagent comprises 30-60mmol/LTris damping fluid, is combined with the latex particle of described monoclonal antibody, 1-5mmol/L tween 20,0.5-3mmol/L sodium azide, 6-13mmol/L sodium ethylene diamine tetracetate and 1-5mmol/L bovine serum albumin.
Above-mentioned hybridoma, called after RBP-2B6, be deposited in Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University preservation center (i.e. China typical culture collection center) on August 28th, 2013, preserving number is CCTCCNo:C2013132.
The secretion output of above-mentioned hybridoma is high, it is secreted the anti-retinol conjugated protein monoclonal antibody obtained and has the advantage such as high-affinity, high specific, Retinal-binding protein detection field can be widely used in, as the preparation field etc. of detection reagent or test set, in specificity, sensitivity and verification and measurement ratio etc., than traditional detection method, there is significant advantage.
Accompanying drawing explanation
Fig. 1 for be restructuring RBP antigen originally when detecting, when to wrap by concentration be 2 μ g/mL, RBP monoclonal antibody titration result figure;
Fig. 2 is detection reagent in embodiment 2 and contrast agents detected result dependency figure.
Embodiment
Mainly in conjunction with the drawings and the specific embodiments the hybridoma and application thereof that can secrete anti-retinol conjugated protein monoclonal antibody are described in further detail below.
The hybridoma secreting anti-retinol conjugated protein monoclonal antibody of present embodiment is deposited in Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University preservation center (CCTCC), and preserving number is CCTCCNo:C2013132.This hybridoma can secrete anti-retinol conjugated protein monoclonal antibody.The monoclonal antibody of this hybridoma and secretion thereof can be applicable to be prepared in Vogan-Neu detection reagent or test set field, thus can be widely used in the detection reagent or test set field preparing diagnosis kidney injury disease or hepatic diseases.
Present embodiment additionally provides a kind of Retinal-binding protein detection test kit, it contains the monoclonal antibody of above-mentioned hybridoma secretion, specifically this monoclonal antibody is wrapped on latex particle, the volume of antigen-antibody binding substances is increased, retinol conjugated protein in calibration object or testing sample can with monoclonal antibody latex particle generation aggregation, form immune complex and produce turbidity, its turbidity height is directly proportional to the retinol conjugated protein in sample when a certain amount of antibody exists.By measuring the absorbance of specific wavelength, the content of retinol conjugated protein in testing sample can be calculated with reference to working curve.Specifically in the present embodiment, this Retinal-binding protein detection test kit comprises the first reagent and the second reagent.Wherein, the first reagent comprises 30-60mmol/LTris damping fluid, 60-95mmol/L polyoxyethylene glycol, 1-5mmol/L sodium azide, 6-13mmol/L sodium ethylene diamine tetracetate and 1-5mmol/L bovine serum albumin; Second reagent comprises 30-60mmol/LTris damping fluid, is combined with the latex particle of described monoclonal antibody, 1-5mmol/L tween 20,0.5-3mmol/L sodium azide, 6-13mmol/L sodium ethylene diamine tetracetate and 1-5mmol/L bovine serum albumin.Detection reagent needed for other can directly as required be prepared in laboratory.
This monoclonal antibody, except being applied to above-mentioned detection agent box, can also be used in other Retinal-binding protein detection test kits or equipment.It will be appreciated by those skilled in the art that, by the monoclonal antibody of present embodiment directly or indirectly in conjunction with other signal group (as magnetic microsphere, horseradish peroxidase etc.), or the monoclonal antibody of present embodiment be can be used in other forms of Retinal-binding protein detection reagent or equipment as coated antibody (such as ELISA).
The secretion output of this hybridoma is high, it is secreted the anti-retinol conjugated protein monoclonal antibody obtained and has the advantage such as high-affinity, high specific, Retinal-binding protein detection field can be widely used in, as the preparation field etc. of detection reagent or test set, in specificity, sensitivity and verification and measurement ratio etc., than traditional detection method, there is significant advantage.
It is below specific embodiment part
The foundation of embodiment 1 hybridoma cell strain and the preparation of anti-RBP monoclonal antibody
1. antigen immune
To recombinate RBP antigen (Shenzhen City Fapon Biotech Co., Ltd produce, article No. BA-PAB-MU0032) equivalent and Freund's complete adjuvant (outward appearance: amber cell suspending liquid; Component: ParaffinOil85%, MannideMonooleate15%, Mycobacteriumsmegmatis1mg/mL) mixing, obtain oil emulsion.By this emulsion with the injected sc BALB/c mouse of 0.2mL (Guangzhou province Experimental Animal Center, 6 week age is female, 5) site, back, first time immunity 14 days pneumoretroperitoneums strengthen immunity (equivalent amount of antigen mixes with Freund's incomplete adjuvant), strengthen immunity after four pins, adopt tail blood and carry out bioactivity, tire and reach fusion requirement.
Merge first 3 days, with same dose antigen abdominal injection supplementary immunization, immunization method is the same.
2. the preparation of hybridoma cell strain
(1) preparation of feeder cell
Feeder cell are made with BALB/c mouse peritoneal macrophage.In fusion first 1 day, BALB/c mouse drew neck to put to death, and 75% alcohol whole body soaks, in super clean bench, with scissors abdominal cut skin under aseptic technique, expose peritonaeum, with syringe Intraperitoneal injection RPMI1640 basic culture solution 5mL, repeatedly rinse, reclaim washing fluid, 1000rpm, centrifugal 5 minutes, stay precipitation, resuspended with RPMI1640 screening and culturing liquid (containing in the RPMI1640 complete culture solution of HAT), adjustment cell concn 1 × 10
5individual/mL, adds 96 orifice plates, 150 μ L/ holes, 37 DEG C, 5%CO
2overnight incubation.
(2) preparation of immune spleen cell
After mouse final immunization three days, aseptically take out spleen, be placed in plate, RPMI1640 basic culture solution rinses once, the nylon wire being put in small beaker grinds filtration, makes cell suspension.Centrifugal, abandon supernatant, RPMI1640 basic culture solution is resuspended, so in triplicate, obtains immune spleen cell, counting.
(3) preparation of myeloma cell
Murine myeloma cell Sp2/0, after 8-anaguanine screening, is cultured to logarithmic phase, gets two large bottles and make cell suspension, centrifugal, abandons supernatant, resuspended with RPMI1640 basic culture solution, obtains myeloma cell in triplicate, counting as a little.
(4) cytogamy and HAT select hybridoma
Myeloma cell is mixed in 1:10 ratio with immune spleen cell, in 50mL plastic cement centrifuge tube, washes 1 time, 1200rpm with RPMI1640 basic culture solution, centrifugal 8 minutes.Abandon supernatant, mixed by cell, the PEG1500 slowly adding 1mL50% merges, and merges the RPMI1640 basic culture solution adding 15mL after 1 minute and stops cytogamy.1000rpm, centrifugal 5 minutes.Abandon supernatant, resuspended gently with the RPMI1640 screening and culturing liquid of 50mL, divide equally in 10 piece of 96 orifice plate, 50 μ L/ holes, 37 DEG C, 5%CO
2cultivate.Be cultured to the 6th day, change HAT nutrient solution (the RPMI1640 complete culture solution containing HAT) twice.
(5) detection of antibody
Natural RBP(SRIPPS is diluted with 0.06MpH9.6 carbonate buffer solution), make its final concentration be 5 μ g/mL.Every hole 0.1mL, adds 96 hole polystyrene plates, and 37 DEG C are spent the night for 2 hours or 4 DEG C.Next day, with the 0.02MpH7.2PBS containing 10% calf serum or 1% skim-milk, 0.15mL/ hole, closes 2 hours for 37 DEG C, for detecting.Merge latter 7th day, get cell conditioned medium 0.1mL in above-mentioned 96 hole check-out consoles, 37 DEG C 30 minutes, sheep anti-mouse igg (the Shenzhen City Fapon Biotech Co., Ltd production of the horseradish peroxidase mark of 2000 times of dilutions is added after washing six times, article No. BA-PAB-MU0032), 37 DEG C 30 minutes the same wash after, every hole adds 100 μ L containing 0.1%(M/V) O-Phenylene Diamine, 0.1%(V/V) hydrogen peroxide, pH5.0 citrate phosphate buffer, 37 DEG C 15 minutes, add dilution heat of sulfuric acid, every hole 50 μ L, surveys 450nm absorption value.RPMI1640 complete culture solution, as negative control, is positive cell hole with measured value and control value Bi≤2.0.
Secretory antibody positive cell hole is cloned with limiting dilution assay on 96 well culture plates with 1 cells/well, and method on positive Kong Yi of screening clones three times, after enlarged culturing continuously, and frozen with the nutrient solution containing 10%DMSO, cell density is 10
6individual/mL.Cytogamy once obtains the cell strain of 1 strain energy stably excreting antibody altogether, called after RBP-2B6(CCTCCNo:C2013132) cell strain.
3. the preparation of monoclonal antibody
Select the BALB/c mouse that 6-8 week is healthy and strong, the pristane of every mouse peritoneal injection 0.5mL; Pneumoretroperitoneum injection 1 × 10 in 10 days
6individual hybridoma.Can produce ascites after inoculating cell 7-10 days, the healthy state of close observation animal and ascites sign, treat that ascites is many as far as possible, and mouse is frequency domain before death, puts to death mouse, suck in test tube with dropper by ascites, a general mouse can obtain 5-10mL ascites.Collect ascites, centrifuging and taking supernatant, is put in-20 DEG C of Refrigerator stores.
Get ascites supernatant, with filter paper filtering after the PBS dilution of 3 times of volumes.The filtrate of gained is added under the flow velocity of 1mL/min one with the protein g affinity chromatography post (purchased from AmershamBiosciences company) of PBS balance.Then the material do not adsorbed by Protein G with the washing of the flow velocity of 1ml/min with PBS is until at OD
280nmunder absorption value reach baseline till.Use glycine elution liquid (pH2.5) wash-out of 0.1M again and reclaim this antibody.The solution 0.1MTris(pH8.8 reclaimed) neutralization, antibody concentration is adjusted to a suitable concentration by ultrafiltration ,-20 DEG C of packing are frozen.
4. titration
Detect with indirect elisa method, above-mentioned Hybridoma Cell Culture supernatant tires 1.22 × 10
3, ascites supernatant tires 1.02 × 10
6.Specific experiment step is as follows:
(1) bag quilt: by restructuring RBP antigen diluent to the 2 μ g/mL of immunity, 100 μ L/ holes add to enzyme plate, and 37 DEG C are spent the night for 2 hours or 4 DEG C;
(2) wash plate 5 times with washing trigger, each every hole note washing lotion 350 μ L, stops 20 seconds; Finally pat dry;
(3) wash away coating buffer with washings, close with confining liquid, every hole 150 μ L, place 1.5-2 hour for 37 DEG C;
(4) wash trigger and wash plate 5 times, each every hole note washing lotion 350 μ L, stops 20 seconds; Finally pat dry;
(5) sample is added: respectively the cells and supernatant and ascites that are diluted to different gradient are added the antigen coated good enzyme plate of above-mentioned restructuring RBP, 100 μ L/ holes, 37 DEG C of reactions, 1 hour (simultaneously doing negative control hole and Positive control wells);
(6) wash trigger and wash plate 5 times, each every hole note washing lotion 350 μ L, stops 20 seconds; Finally pat dry;
(7) add the sheep anti-mouse igg (diluting 6000 times with confining liquid) of horseradish peroxidase mark, 100 μ L/ holes, 37 DEG C are reacted 1 hour;
(8) wash trigger and wash plate 5 times, each every hole note washing lotion 350 μ L, stops 20 seconds; Finally pat dry;
(9) add nitrite ion TMB: instantly to join, 100 μ L/ holes, 37 DEG C of lucifuges react 30 minutes;
(10) termination reaction: the sulfuric acid adding 2M in each reacting hole, 50 μ L/ holes;
(11) microplate reader reading: 450nm, 630nm wavelength measures, it is 1.22 × 10 that the final reaction measuring this Hybridoma Cell Culture supernatant and restructuring retinol conjugated protein antigen is tired
3, it is 1.02 × 10 that ascites antibody is tired with restructuring retinol conjugated protein antigen-reactive
6.
By ELISA method, measure absorbancy at 450nm, 630nm wavelength, use Subclass of antibody identification kit, identify RBP monoclonal antibody class, subclass, table 1 is qualification result.
Table 1RBP monoclonal antibody type identification result
Fig. 1 for be restructuring RBP antigen originally when detecting, when to wrap by concentration be 2 μ g/mL, RBP monoclonal antibody titration result figure.
5. Identification of Monoclonal Antibodies
Avidity: the monoclonal antibody of above-mentioned RBP-2B6 hybridoma secretion detects with ELISA method, and its affinity constant (Kaff) is 9.8 × 10
8.Be below the concrete process measuring the monoclonal antibody avidity of RBP-2B6 hybridoma secretion:
The concentration of monoclonal antibody after employing ultraviolet absorption method mensuration purifying, that is:
Protein concentration (mg/mL)=1.45 × OD280nm ﹣, 0.74 × OD260nm
Adopt ELISA method to calculate affinity costant, step is as follows: by 4 μ g/mL, 2 μ g/mL and 1 μ g/mLRBP recombinant antigen wrapper sheet enzyme plate respectively, and add concentration known and carry out the anti-RBP monoclonal antibody of doubling dilution, other is with above-mentioned titration step.Last with the logarithmic value of each monoclonal antibody different concns for X-coordinate, with its corresponding 450nm, 630nm wavelength absorbance is ordinate zou, draw out three response curves of different concns antigen, getting the numerical value that each curve top is tending towards flat sections is OD-100, finds monoclonal antibody concentration [Ab] t of its OD-50 point, therefore can obtain [Ab] t, [Ab '] t and [Ab 〞] t tri-values, by derivation formula calculating K aff values such as following Beatty:
When envelope antigen concentration is 2 μ g/mL:
Kaff1=1/2（2[Ab′]t-[Ab]t）
Or Kaff2=1/2(2 [Ab 〞] t-[Ab '] t)
During 1 μ g/mL: Kaff3=3/2(4 [Ab 〞] t-[Ab] t)
Such monoclonal antibody measures at every turn and can obtain three Kaff values, and mean value i.e. its affinity constant Kaff.
Embodiment 2RBP latex immunoturbidimetry detection kit
1, the main agents of the detection kit of the present embodiment is formulated as follows:
(1) reagent R1 comprises: 45mmol/LTris damping fluid, 85mmol/L polyoxyethylene glycol, 5mmol/L sodium azide, 8.5mmol/L sodium ethylene diamine tetracetate and 5mmol/L bovine serum albumin.
(2) reagent R2 comprises: 45mmol/LTris damping fluid, be combined with the latex particle 5%(v/v of RBP monoclonal antibody), 5mmol/L tween 20,3mmol/L sodium azide, 8.5mmol/L sodium ethylene diamine tetracetate, 5mmol/L bovine serum albumin.
(3) calibration object: be serum matrix for detecting the calibration object of serum, concentration is respectively 130mg/L, 100mg/L, 50mg/L, 25mg/L, 12.5mg/L, 0mg/L, or the content of similar ratio; Be urine matrix for detecting the calibration object of urine, concentration is respectively 4mg/L, 2mg/L, 1mg/L, 0.5mg/L, 0.25mg/L, 0mg/L, or the content of similar ratio.
2 testing processes
(1) reagent prepares, and reagent R1 comprises 45mmol/LTris damping fluid, 85mmol/L polyoxyethylene glycol, 5mmol/L sodium azide, 8.5mmol/L sodium ethylene diamine tetracetate and 5mmol/L bovine serum albumin; Reagent R2 comprises 45mmol/LTris damping fluid, is combined with the latex particle of RBP monoclonal antibody, 5mmol/L tween 20,3mmol/L sodium azide, 8.5mmol/L sodium ethylene diamine tetracetate and 5mmol/L bovine serum albumin; RBP calibration object.
(2) detecting step (completing in automatic clinical chemistry analyzer)
The RBP detection kit that the present embodiment describes.Be applicable to various types of full automatic biochemical apparatus, for Hitachi 7170 full automatic biochemical apparatus, its operation is as follows, adopt Two point end assay, concrete steps are as follows, 120 μ L reagent R1 add 3 μ L samples after 37 DEG C of 5min, add 120 μ L reagent R2, namely start reading, read another point after reaction 5min; Determined wavelength predominant wavelength 570nm, commplementary wave length 800nm.By the relation that RBP concentration in calibration object and absorbancy change, set up working curve, measure the absorbance in measuring samples, the RBP concentration in sample can be calculated with reference to working curve.
The application of embodiment 3RBP latex immunoturbidimetry detection kit
Using the reagent of this law invention, adopt Hitachi 7170 full automatic biochemical apparatus, is 110mg/L to interpolation concentration, 60mg/L, 15mg/LRBP serum sample and 3.5mg/L, 2mg/L, 0.2mg/LRBP urine specimen detect, each test sample 5 times, calculate its mean value, standard deviation, the variation coefficient, to investigate the stability result of test kit in table 2, table 3.Meanwhile, use the reagent controls of this law invention certain commercially available reagent external, adopt Hitachi 7170 full automatic biochemical apparatus, measure 50 parts of serum samples simultaneously, carry out correlation analysis to measured value, linear equation is Y=0.98733X-2.89949, coefficient R=0.98602, the results are shown in Figure 2.
Table 2 serum sample adds recovery test
Table 3 urine specimen adds recovery test
This kit results accuracy is strong, reagent stability good, easy to use, be convenient to large-scale promotion.
The above embodiment only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but therefore can not be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (8)
1. can secrete a hybridoma for anti-retinol conjugated protein monoclonal antibody, preserving number is CCTCCNo:C2013132.
2. can secrete the application of hybridoma in preparation diagnosis kidney injury disease or the detection reagent of hepatic diseases or the test set of manufacture diagnosis kidney injury disease or hepatic diseases of anti-retinol conjugated protein monoclonal antibody as claimed in claim 1.
3. can secrete the hybridoma of anti-retinol conjugated protein monoclonal antibody as claimed in claim 1 in the application preparing Retinal-binding protein detection reagent or manufacture in the test set of retinol conjugated protein.
4. a monoclonal antibody of being secreted by the hybridoma can secreting anti-retinol conjugated protein monoclonal antibody as claimed in claim 1.
5. the application of monoclonal antibody as claimed in claim 4 in preparation diagnosis kidney injury disease or the detection reagent of hepatic diseases or the test set of manufacture diagnosis kidney injury disease or hepatic diseases.
6. monoclonal antibody as claimed in claim 4 is in the application preparing Retinal-binding protein detection reagent or manufacture in the test set of retinol conjugated protein.
7. a Retinal-binding protein detection test kit, is characterized in that, comprises monoclonal antibody as claimed in claim 4.
8. Retinal-binding protein detection test kit as claimed in claim 7, it is characterized in that, comprise the first reagent and the second reagent, wherein, described first reagent comprises 30-60mmol/LTris damping fluid, 60-95mmol/L polyoxyethylene glycol, 1-5mmol/L sodium azide, 6-13mmol/L sodium ethylene diamine tetracetate and 1-5mmol/L bovine serum albumin; Described second reagent comprises 30-60mmol/LTris damping fluid, is combined with the latex particle of described monoclonal antibody, 1-5mmol/L tween 20,0.5-3mmol/L sodium azide, 6-13mmol/L sodium ethylene diamine tetracetate and 1-5mmol/L bovine serum albumin.
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| CN102628867A (en) * | 2011-12-30 | 2012-08-08 | 北京九强生物技术股份有限公司 | Double antibody latex enhanced retinol binding protein detection kit |
| CN102841210A (en) * | 2012-08-30 | 2012-12-26 | 谢兵 | Retinol detection kit and preparation method thereof |
| CN102944679A (en) * | 2012-11-15 | 2013-02-27 | 北京康美天鸿生物科技有限公司 | Kit for performing retinol binding protein detection by using latex turbidimetry |
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| CN102628867A (en) * | 2011-12-30 | 2012-08-08 | 北京九强生物技术股份有限公司 | Double antibody latex enhanced retinol binding protein detection kit |
| CN102841210A (en) * | 2012-08-30 | 2012-12-26 | 谢兵 | Retinol detection kit and preparation method thereof |
| CN102944679A (en) * | 2012-11-15 | 2013-02-27 | 北京康美天鸿生物科技有限公司 | Kit for performing retinol binding protein detection by using latex turbidimetry |
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