CN1116548A - Polyclonal antibody, preparing process and use thereof - Google Patents
Polyclonal antibody, preparing process and use thereof Download PDFInfo
- Publication number
- CN1116548A CN1116548A CN 95111630 CN95111630A CN1116548A CN 1116548 A CN1116548 A CN 1116548A CN 95111630 CN95111630 CN 95111630 CN 95111630 A CN95111630 A CN 95111630A CN 1116548 A CN1116548 A CN 1116548A
- Authority
- CN
- China
- Prior art keywords
- polyclonal antibody
- kinds
- chemical compound
- hapten
- ethyl ester
- 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.)
- Granted
Links
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Abstract
The multi-clon antibody for catalyzing the hydrolysis of naproxen ester, with unique selectivity of chiral naproxen ester and for resolution of (S)-naproxen is prepared with monomethyl or monoethyl 1-(6-hydroxy-beta-naphthyl)-ethyl-phosphonate as semi-antibody and butanedioic or glutaric anhydride as bridge by linking one carboxyl with 6-hydroxy in semi-antibody to form ester, linking another carboxyl with carrier albumen, e.g. BSA (bovine serum albumin) or keyhole-limpethemocyanin, to form semi-antibody-carrier albumen system, animal immune, separation of antigen serum and purifying.
Description
The present invention relates to a kind of polyclonal antibody, preparation method and its usage.This polyclonal antibody is by 1-(6-hydroxy-beta-naphthyl)-ethylphosphonic acid mono ethyl ester hapten, connect the antigen system that is suitable for animal immune that forms by dibasic acid anhydride as bridge and carrier protein, behind animal immune, obtain polyclonal antibody again, have the esterolytic ability of catalysis naproxen.
A great discovery catalytic antibody (abzyme) (Tramontano A, Janda K.D.Lerner R.A.Catalytic Antibodies.Science, 1986,234:1566-1570 in the eighties chemistry and the field of immunology; Pollack S.J.Jacobs J.W., Schultz P.G.Selective Chemical Catalysis by an Antibody, Science, 1986,234:1570-1573), the extremely multiformity of antibody and the characteristics such as huge catalytic capability of enzyme are combined, for a brand-new direction has been opened up in traditional chemistry and immunology research.Utilize through the special organic molecule that designs and synthesizes as hapten, connect behind the carrier protein immune animal and through hybridoma technique, from screening and isolating monoclonal antibody, successfully acquisition has the antibody of catalysis.So far existing more than 50 reaction can with catalytic antibody come catalysis (Schultz P.G., Lerner R.A., Benkovic S.J.Catalytic Anti-bodies.Chem.Eng.News 1990, May 28, P20-40; Ikeda S., the progressive recently と of Janda K.D.Catalytic Antibody (catalyst antibody) uses To つ ぃ て. Synthetic Organic Chemistry association will, 1993,51:284-297), to some reaction (as the Diels-Alder reaction), there is not their enzyme of energy catalysis in nature, can carry out catalysis (Hilvert D.et al.Antibody Catalysis of a Diels-Alder Reaction.Am.Chem.Soc.1989,111:9261-9262 with antibody yet; Braisted A.C., Schultz P.G. An Anti-body-CatalyzedBimolecular Diels-Alder Reaction. J.Am.Chem.Soc.1990,112:7430-7431).To very few (the Gallacher D.et all.A polyclonal antibody preparation with Michaelian catalytic proper-ties.Biochem.J.1991 of report that comes catalyse organic reaction with polyclonal antibody, 279,871-881.Polyclonal antibody-catalysed amidehydrolysis.ibid 1992,284,675-680).We have reported 1-(6-hydroxy-beta-naphthyl)-ethylphosphonic acid mono ethyl ester as hapten, but do not see the report of polyclonal antibody to chiral Recognition so far.Therefore, design has specific catalytic antibody, and at chemistry, biology, fields such as medical science will show huge application potential undoubtedly.Naproxen is a kind of very important non-steroidal anti-inflammatory analgesics, and the drug effect of its S configuration is 38 times of R configuration, and research splits the naproxen that obtains the S configuration with antibody catalysis hydrolysis naproxen ethyl ester, has important significance for theories and potential using value.
The purpose of this invention is to provide a kind of polyclonal antibody.
Another object of the present invention provides a kind of method for preparing this polyclonal antibody.
A further object of the invention provides the purposes of this polyclonal antibody.
A kind of polyclonal antibody of the present invention is to be hydrolysis substrate with the naproxen ester, designing its phosphonate ester (4) is transition state analogs, with 1-(6-hydroxy-beta-naphthyl)-ethylphosphonic acid mono-methyl or mono ethyl ester (4) is hapten, with succinic anhydride or glutaric anhydride is bridge, a carboxyl and haptenic 6-hydroxyl connect, obtain being fit to the hapten-carrier albumen system (5) of animal immune
R=Me or Et in the formula, R
1=BSA or KLH, R
2=(CH
2)
2Or (CH
2)
3Described carrier protein is bovine serum albumin or key Kong Korea Spro hemocyanin, can abbreviate BSA or KLH respectively as.With hapten-carrier albumen system (5) immune animal, tell serum, the purified immunoglobulin (IgG) that obtains, polyclonal antibody promptly of the present invention after getting blood.
In other words polyclone of the present invention be a kind of be the polyclonal antibody that hapten makes with 1-(6-hydroxy-beta-naphthyl)-ethane phosphonic acid monoesters or ethyl ester (4).Also we can say polyclonal antibody of the present invention be a kind of with hapten one carrier protein system (5) through immune animal, get blood system and go out whole serum, separation and purification obtains polyclonal antibody.
Polyclonal antibody of the present invention adopts pH when being 7.3 phosphate buffer, and capillary electrophoresis detects shows the homogeneous product.
Polyclonal antibody molecular weight of the present invention reaches 150000 approximately, whole molecule is made up of four polypeptide chains, wherein two identical long-chains are heavy chain, molecular weight respectively is about 50000, two identical light chains, and molecular weight is about 25000, article two, heavy chain mat disulfide bond is tied, be the Y font, two light chains are connected the both sides of Y word again by disulfide bond, and whole antibody molecule is symmetrical structure.
Preparation method of polyclonal antibody of the present invention, be to be raw material with 1-(6-hydroxy-beta-naphthyl)-ethylphosphonic acid mono-methyl or ethyl ester (4) hapten, make bridge with succinic anhydride or glutaric anhydride, make a carboxyl and 6-hydroxyl be connected into ester, another carboxyl connects with carrier protein, described carrier protein is bovine serum albumin (BSA) or key hole hemocyanin (KLH), obtains being suitable for the antibody of animal immune.
Wherein R=Me or Et, R
1=BSA or KLH, R
2=(CH
2)
2Or (CH
2)
3Specifically at room temperature with polar solvent in, (simple DMAP) is catalyst with right-dimethylamino naphthyridine, chemical compound (4) and succinic anhydride or glutaric anhydride reaction promptly got chemical compound (6) in 1-20 hour, and wherein the mole ratio of chemical compound (4), succinic anhydride or glutaric anhydride and catalyst is 1: 1-10: 0.1-2.The recommendation ratio is 1: 1-5: 0.8-1.5.Described solvent is pyridine preferably.
In polar solvent, chemical compound (6) can obtain hapten-carrier albumen (5) in 0.5-10 hour with carrier protein BSA or KLH reaction.Usually the mole ratio of chemical compound (6) and carrier is 1: 0.001-0.1, with 1: 0.001-0.01 is for well.Reaction can at room temperature be carried out.Add 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide condensing agent and will help adding the carrying out of fast response, the mole ratio of chemical compound (6) and condensing agent is 1: 0.5-10.Usually add reaction system after carrier protein is dissolved in phosphate buffer.
The hapten-carrier albumen (5) of above-mentioned reaction gained can be removed micromolecular unreacted hapten and condensing agent with the semipermeable membrane dialysis.
Hapten-carrier albumen (5) is carried out animal immune, get blood and isolate anti-whole serum, get rough polyclonal antibody with the saturated ammonium sulfate solution precipitation, specifically, hapten-carrier albumen system and Freund adjuvant (Freund ' s adjurant) equal-volume is mixed, make the emulsifying vaccine, carry out animal immune, make initial immunity at the position subcutaneous injection vaccines such as neck, the back of the body and vola of animals such as mice, rabbit, rat or Cavia porcellus.When immune once more, the Freund adjuvant that replaces initial immunity with incomplete Freund's adjuvant is made vaccine with immune animal.Be 10-30 days twice immune blanking time.Last immunity was taken out blood in back 7 days, isolated anti-whole serum, and the saturated ammonium sulfate solution precipitation with 40% gets thick polyclonal antibody.Can separate with the DE-52 ion-exchange chromatography and remove other serum albumin, get pure polyclonal antibody, also available freeze-drying obtains pure polyclonal antibody powder, the energy long preservation, and capillary electrophoresis shows homogeneous.
The immune effect of animal immune proof rabbit is best, uses enzyme linked immunosorbent assay (ECISA) to measure when haptenic antibody titer (O.D. value) dilutes 6400 times in the serum, and its antibody titer still is 3.4 times of blank group.
Polyclonal antibody of the present invention can general methyl ester of catalysis naphthalene or ethyl ester hydrolysis generation naproxen.When catalysis raceme naproxen ethyl ester, its catalytic hydrolysis reaction kinetics meets the Michaelis-Menten rate equation.Fig. 1 is the concentration of substrate time curve of hydrolysis, is expressed as contrast among Fig. 1 and do not add the catalysis polyclonal antibody,
*For adding the result of catalysis polyclonal antibody.Fig. 2 is the inverse of concentration of substrate and the relation curve of its initial velocity inverse, obtains from the gained data computation: Vmax=17.4 ± 0.8 μ mol/L.min, K
M=477 ± 13 μ mol/LKcat=2.26 ± 0.10min
-1, kcat/Kuncat=1240, Vmax/[E
0]=135S
-1. show the general ethyl ester hydrolysis of polyclonal antibody energy catalysis naphthalene of the present invention, quicken 1240 times.Polyclonal antibody of the present invention finds that respectively to the S-naproxen ethyl ester and the R-naproxen ethyl ester catalyzing hydrolysis of optical activity the concentration of S-isomer remains unchanged, and the concentration of R-isomer is in continuous reduction, and about 4-8 hour, the energy complete hydrolysis was fallen the ester of R-isomer.The results are shown among Fig. 3 and Fig. 4.Fig. 3 represents the relation in concentration of substrate and response time, wherein,
, ■ and Q, , ● represent S-naproxen ethyl ester and R-naproxen ethyl ester respectively,, zero and ■, ● expression does not add and adds the result of polyclonal antibody respectively.
, is the corresponding polyclonal result of non-specificity who adds the matched group rabbit.Fig. 4 is the inverse of R-naproxen ethyl ester concentration of substrate and the relation curve of its initial velocity inverse, obtains K from straight slope
M=202 μ M obtain Vmax=417 μ M.min from the straight line intercept
-1, Kcat=0.0496s
-1/ uncat=1285. is that polyclonal antibody of the present invention can make the hydrolysis speed of R-naproxen ethyl ester increase by 1285 times.
Polyclonal antibody of the present invention not only preparation method is easy, but product long preservation, and can be used for splitting the naproxen ethyl ester of DL, after enantioselective hydrolysis falls the R-configuration, the acid that the R-configuration is removed in available diluted alkaline washing, with reactant mixture saponification under alkali condition, the separable naproxen that goes out the S-configuration.
To help further to understand the present invention by following embodiment, but not limit content of the present invention.The preparation of embodiment 1 chemical compound (6)
Chemical compound (4) (0.8mmol) is dissolved in the 2mL pyridine, and adding succinic anhydride or glutaric anhydride (1.4mmol) and DMAP (100mg, 0.8mmol).At room temperature stir and spend the night, the thin layer chromatography detection reaction is complete.Silica gel column chromatography separates, and with petroleum ether flush away pyridine, uses ethyl acetate drip washing then earlier, the mixed solvent drip washing of reuse ethyl acetate dehydrated alcohol (1: 2) afterwards, concentrate white product 160-170mg.
Work as R=Et, when adopting succinic anhydride, product (9)
1H NMR:
7.86(s，1H，ArH)，7.83(d，J＝8.7H
z，1H，ArH)，7.73(d，J＝8.5H
z，
1H，ArH)，7.64(d，J＝8.5H
z，1H，ArH)，7.25(d，J＝2.16H
z，1H，
ArH)，7.21(dd，J＝8.7，2.16H
z，1H，ArH)，6.85(d，J＝7.4H
z，1H，
OH)，4.02(q，J＝1.5H
z，2H，POCH
2CH
3)，3.62(q，J＝7.3H
z，2H，
ArOCOCH
2CH
2COOH)，3.37(m，1H，ArCHCH
3)，3.25(q，J＝7.3
H
z，2H，ArOCOCH
2CH
2COOH)，1.77(dd，J＝7.2，16.6H
z，3H，
ArCHCH
3)，1.44(t，J＝1.5H
z，3H，POCH
2CH
3).
MS:381 (M+1), the preparation of 335,319,303,280,265,171,102. embodiment 2 chemical compounds (5)
BSA or KLH 10mg are dissolved in the 3.75mL phosphate buffer, the N that under stirring at room, adds chemical compound (6) 30mg, dinethylformamide 1.25mL solution and 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide condensing agent 10-30mg, reacted 1 hour, remove micromolecule with the semipermeable membrane dialysis, get hapten-carrier albumen (5).Embodiment 3 Polyclonal Antibody Preparation
Hapten-carrier albumen (5) and Freund adjuvant equal-volume is mixed, make emulsive vaccine, make initial immunity at the position subcutaneous injection vaccines such as neck, the back of the body and vola of animal.When immune again, the Freund adjuvant when replacing initial immunity with incomplete Freund's adjuvant is made vaccine with immune animal.Twice immune about 10-30 days blanking time.Blood was got in last immunity in back 7 days, and blood is placed and spent the night, and allows it condense, and through the centrifugal clot of removing, serum is partly used 40% saturated ammonium sulfate solution precipitation.The centrifugal supernatant that goes, precipitation is analysed separation with the DE-52 resinbed and is removed other serum albumin with the least possible phosphate buffer dissolving after dialysing, obtain immunoglobulin IgG, capillary electrophoresis show the homogeneous product (phosphate buffer, pH7.3), i.e. polyclonal antibody.
Measure the antibody titer of antihapten in the above-mentioned serum with enzyme linked immunological absorption method (ELISA), adopted be different from hapten-carrier that the carrier protein for preparing vaccine forms as envelope antigen or with blockade, in and the method (Ji Yongyong of anti-carrier protein antibody in the antiserum, China's Journal of Immunology, 7:290-293,1991), measurement result is as follows:
Antibody titer (the OD of antihapten in the immune serum
450)
The kinetic determination of embodiment 4 catalysis racemization naproxen ethyl ester hydrolysis
| Animal | Numbering | Serum diluting multiple | Antibody titer (OD 450mm) |
| Mice | ????1 ????2 ????3 ????4 ????5 ????6 | ????＜50 ????＜50 ????＜50 ????＜50 ????＜50 ????＜50 | ????0.397 ????0.409 ????0.425 ????0.331 ????0.425 ????0.425 |
| The contrast contrast | ????50 ????200 | ????0.335 ????0.214 | |
| Rabbit | ????1 ????2 | ????＞6400 ????＞6400 | ????0.660 ????0.668 |
| Contrast | ????6400 | ????0.200 | |
| Rat | 123 contrast contrasts | ????50 ????＜50 ????800 ????50 ????800 | ????0.452 ????0.324 ????0.333 ????0.332 ????0.181 |
| Cavia porcellus | ????1 ????2 ????3 | ????1600 ????100 ????100 | ????0.266 ????0.407 ????0.327 |
| Contrast | ????1600 | ????0.154 |
Hydrolysising condition: at 50mmol/L Tris-HCl (Tri(Hydroxymethyl) Amino Methane Hydrochloride), 136mmol/L NaCl, pH=8.2, under 37 ℃ of conditions, the ratio of racemization naproxen ethyl ester substrate and polyclonal antibody is 50: 1, the blank condition is identical.
Analysis condition: with HPLC C
18Reversed-phase column is followed the tracks of hydrolysis, and the detection wavelength is 240mm, and mobile phase is acetonitrile: water=70: 30, interior mark are 6-methoxyl group-β-naphthalene methanol.
The result is plotted in Fig. 1 and Fig. 2.Embodiment 5 catalysis R-and S-configuration naproxen ethyl ester hydrolysis kinetic determination
Hydrolysising condition: at 50mmol/L Tris-HCl (Tri(Hydroxymethyl) Amino Methane Hydrochloride), 100mmol/L NaCl, pH=8.2, under 37 ℃ of conditions, R or S-configuration ethyl ester respectively are 30mol, polyclonal antibody 0.7 μ mol, the blank condition is identical.Analysis condition is with embodiment 4, and the result is plotted in Fig. 3 and Fig. 4.
Claims (10)
1 one kinds of polyclonal antibodies is characterized in that with molecular formula being
Chemical compound as hapten, preparation hapten-carrier albumen through animal immune, is got blood, tells anti-whole serum, purification makes, wherein R=Me or Et, R
1=BSA or KLH, R
2=(CH
2)
2Or (CH
2)
3
2 one kinds of polyclonal antibodies as claimed in claim 1 is characterized in that with pH being that 7.3 phosphate buffer capillary electrophoresis detects and shows the homogeneous product.
3 one kinds of Polyclonal Antibody Preparation methods as claimed in claim 1 is characterized in that recording with following step:
(1) in room temperature and polar solvent, right-dimethylamino naphthyridine is a catalyst, 1-(6-hydroxy-beta-naphthyl)-ethylphosphonic acid mono-methyl or mono ethyl ester (4) and (R
2CO)
2The O reaction got chemical compound (6) in 1-20 hour,
Wherein R=Me or Et, R
1=OH, R
2=(CH
2)
2Or (CH
2)
3, chemical compound (4), (R
2CO)
2O and catalyst mole ratio are 1: 1-10: 0.1-2,
(2) in polar solvent, chemical compound (6) and carrier protein BSA or KLH at room temperature reacted 0.5-10 hour to such an extent that molecular formula is
Hapten-carrier albumen (5), R wherein
1=BSA or KLH, R=Me or Et, R
2=(CH
2)
2Or (CH
2)
3, the mole ratio of chemical compound (6) and carrier protein is 1: 0.001-0.1,
(3) hapten-carrier albumen (5) and Freund adjuvant equal-volume are made the emulsifying vaccine after mixed, carry out the animal initial immunity, replace Freund adjuvant with incomplete Freund's adjuvant when immune once more and make the vaccine immunity animal, twice immune interval 10-30 days, blood was taken out in last immunity in back 7 days, tell anti-whole serum, the saturated ammonium sulfate solution precipitation with 40% gets thick polyclonal antibody.
4 one kinds of Polyclonal Antibody Preparation methods as claimed in claim 3 is characterized in that when preparation chemical compound (6) chemical compound (4), (R
2CO)
2The mole ratio of O and catalyst is 1: 1-5: 0.8-1.5.
5 one kinds of Polyclonal Antibody Preparation methods as claimed in claim 3 is characterized in that solvent is a pyridine when preparation chemical compound (6).
6 one kinds of Polyclonal Antibody Preparation methods as claimed in claim 3, it is characterized in that also adding 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide condensing agent when preparation hapten-carrier albumen (5), the mole ratio of chemical compound (6) and condensing agent is 1: 0.5-10.
The preparation method of antibody falls in 7 one kinds of many grams as claimed in claim 3, when it is characterized in that animal immune, recommends animal to adopt rabbit.
8 one kinds of Polyclonal Antibody Preparation methods as claimed in claim 3 is characterized in that thick polyclonal antibody is removed other serum albumin with the separation of DE52 ion exchange resin.
9 one kinds of purposes as claim 1,2,3,4,5,6,7 or 8 described polyclonal antibodies is characterized in that catalyzing hydrolysis naproxen methylester or ethyl ester.
10 1 kinds of purposes as claim 1,2,3,4,5,6,7 or 8 described polyclonal antibodies is characterized in that catalyzing hydrolysis R configuration naproxen methylester or ethyl ester, split from the naproxen methylester of racemization or ethyl ester and obtain S configuration naproxen methylester or ethyl ester.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN95111630A CN1055933C (en) | 1995-05-09 | 1995-05-09 | Polyclonal antibody, preparing process and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN95111630A CN1055933C (en) | 1995-05-09 | 1995-05-09 | Polyclonal antibody, preparing process and use thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1116548A true CN1116548A (en) | 1996-02-14 |
| CN1055933C CN1055933C (en) | 2000-08-30 |
Family
ID=5078897
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN95111630A Expired - Fee Related CN1055933C (en) | 1995-05-09 | 1995-05-09 | Polyclonal antibody, preparing process and use thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1055933C (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100427941C (en) * | 2006-04-06 | 2008-10-22 | 复旦大学 | Chiral sensor based on ox seralbumin and preparing process thereof |
| CN100439914C (en) * | 2006-04-06 | 2008-12-03 | 复旦大学 | Method for detecting chiral isomer |
| CN100482643C (en) * | 2007-01-19 | 2009-04-29 | 中国科学院广州生物医药与健康研究院 | Semicarbazide derivative, monoclonal antibody thereof and application |
| CN101161681B (en) * | 2007-11-01 | 2010-12-15 | 江南大学 | Method for preparing double-flumethasone artificial antigen |
| CN101161679B (en) * | 2007-11-01 | 2011-07-20 | 江南大学 | Method for preparing dexamethasone artificial antigen |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SE8802575D0 (en) * | 1988-07-08 | 1988-07-08 | Wallac Oy | TERPYRIDINE DERIVATIVES |
| US5290551A (en) * | 1990-05-08 | 1994-03-01 | Thomas Jefferson University | Treatment of melanoma with a vaccine comprising irradiated autologous melanoma tumor cells conjugated to a hapten |
| DE4135542A1 (en) * | 1991-10-28 | 1993-04-29 | Boehringer Mannheim Gmbh | STORAGE PROTEIN SOLUTIONS |
-
1995
- 1995-05-09 CN CN95111630A patent/CN1055933C/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100427941C (en) * | 2006-04-06 | 2008-10-22 | 复旦大学 | Chiral sensor based on ox seralbumin and preparing process thereof |
| CN100439914C (en) * | 2006-04-06 | 2008-12-03 | 复旦大学 | Method for detecting chiral isomer |
| CN100482643C (en) * | 2007-01-19 | 2009-04-29 | 中国科学院广州生物医药与健康研究院 | Semicarbazide derivative, monoclonal antibody thereof and application |
| CN101161681B (en) * | 2007-11-01 | 2010-12-15 | 江南大学 | Method for preparing double-flumethasone artificial antigen |
| CN101161679B (en) * | 2007-11-01 | 2011-07-20 | 江南大学 | Method for preparing dexamethasone artificial antigen |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1055933C (en) | 2000-08-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA1340511C (en) | Method of catalyzing chemical reactions using monoclonal antibodies | |
| Li et al. | Antibody catalyzed cationic cyclization | |
| US5716836A (en) | Anti-sulfated tyrosine antibody specific for sulfated tyrosine, process for producing the same, and hybridoma capable of producing anti-sulfated tyrosine monoclonal antibody specific for sulfated tyrosine | |
| CN1055933C (en) | Polyclonal antibody, preparing process and use thereof | |
| CN110196334A (en) | Thrombomodulin chemiluminescence immunoassay immue quantitative detection reagent box and preparation method thereof | |
| JPH05203652A (en) | Antibody enzyme immunoassay | |
| FI70726C (en) | HOMOGENEAL IMMUNOANALYSFOERFARANDE MED FLAVINADENINNUCLOTID SOM MAERKESKONPONENT OCH DETEKTERING AV DET MED APOGLUCOSOXIDAS | |
| JPH02500162A (en) | Antibody binding moieties exhibiting stereoselective synthase activity and methods of using the same | |
| US5047330A (en) | Compound labelled by the acetyl cholinesterase of Electrophorus electricus, its preparation process and its use as a tracer or marker in enzymoimmunological determinations | |
| US5236836A (en) | Autoantibodies which enhance the rate of a chemical reaction | |
| KR100998419B1 (en) | hapten compound and antibody | |
| CN109338477A (en) | Based on the DNA encoding library of molecules and its synthetic method of annular template and application | |
| CN111377888B (en) | Rhododendrin mollis toxin III hapten as well as preparation method and application thereof | |
| CN1104216A (en) | Artifical antigen preparation-hapten and protein cross-link method | |
| US20040059096A1 (en) | Anti-ciguatoxin monoclonal antibody | |
| CN102807614A (en) | Hapten synthesis method of pyrethroids alpha-carboxy ethyl sulfur formyl deltamethrin | |
| Hilvert et al. | [18] Antibody catalysis of concerted, carbon carbon bond-forming reactions | |
| ITMI941499A1 (en) | DETERMINATION AND INSULATION OF TAXOLO AND 10-DESACETYL-BACATINA III AND ANALOGUES THROUGH ANTIBODIES | |
| CN106526188A (en) | Detection kit for salbutamol in foods | |
| Liu et al. | An enzyme-like polyclonal antibody capable of catalyzing ester hydrolysis | |
| CN1055122C (en) | Structural method of anti-body albumen A-DNA probe | |
| EP0051593A1 (en) | Isolation of carbohydrates from cell tissue | |
| Krief et al. | Recognition of β-Ketoalcohol-derived Haptens by Tailor-made Antibodies | |
| KR100804022B1 (en) | Bisazacrownethercalix[4]arene linkers and method for synthesizing the same | |
| JP2024006375A (en) | Biotin labeling agent, biotin-labeled proteins, and immunoassay reagents containing biotin labeling agent |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |