IE63385B1 - A method for the selective immunological determination of intact procollagen peptide (Type III) and procollagen (Type III) in body fluids and means for carrying it out - Google Patents
A method for the selective immunological determination of intact procollagen peptide (Type III) and procollagen (Type III) in body fluids and means for carrying it outInfo
- Publication number
- IE63385B1 IE63385B1 IE128988A IE128988A IE63385B1 IE 63385 B1 IE63385 B1 IE 63385B1 IE 128988 A IE128988 A IE 128988A IE 128988 A IE128988 A IE 128988A IE 63385 B1 IE63385 B1 IE 63385B1
- Authority
- IE
- Ireland
- Prior art keywords
- type iii
- peptide
- procollagen
- antibodies
- bound
- Prior art date
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 48
- 108010050808 Procollagen Proteins 0.000 title claims abstract description 36
- 238000000034 method Methods 0.000 title claims description 22
- 230000001900 immune effect Effects 0.000 title claims description 7
- 210000001124 body fluid Anatomy 0.000 title abstract description 10
- 239000010839 body fluid Substances 0.000 title abstract description 10
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 11
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 11
- 230000002163 immunogen Effects 0.000 claims abstract description 8
- 241001465754 Metazoa Species 0.000 claims abstract description 7
- 239000000427 antigen Substances 0.000 claims description 12
- 102000036639 antigens Human genes 0.000 claims description 12
- 108091007433 antigens Proteins 0.000 claims description 12
- 210000002966 serum Anatomy 0.000 claims description 11
- 230000003053 immunization Effects 0.000 claims description 8
- 238000002649 immunization Methods 0.000 claims description 8
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 5
- 108060003552 hemocyanin Proteins 0.000 claims description 5
- 102000009027 Albumins Human genes 0.000 claims description 3
- 108010088751 Albumins Proteins 0.000 claims description 3
- 241000283707 Capra Species 0.000 claims description 3
- 241001494479 Pecora Species 0.000 claims description 3
- 108010039918 Polylysine Proteins 0.000 claims description 3
- 241000283984 Rodentia Species 0.000 claims description 3
- 229920000656 polylysine Polymers 0.000 claims description 3
- 102000001187 Collagen Type III Human genes 0.000 abstract description 2
- 108010069502 Collagen Type III Proteins 0.000 abstract description 2
- 238000010790 dilution Methods 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 241000894007 species Species 0.000 description 5
- 239000000047 product Substances 0.000 description 4
- 241000700159 Rattus Species 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 238000003127 radioimmunoassay Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 108010035532 Collagen Proteins 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 238000010908 decantation Methods 0.000 description 2
- 238000003018 immunoassay Methods 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000000159 protein binding assay Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- WNWVKZTYMQWFHE-UHFFFAOYSA-N 4-ethylmorpholine Chemical group [CH2]CN1CCOCC1 WNWVKZTYMQWFHE-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 241001474728 Satyrodes eurydice Species 0.000 description 1
- 239000012506 Sephacryl® Substances 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 239000012888 bovine serum Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- VDQQXEISLMTGAB-UHFFFAOYSA-N chloramine T Chemical compound [Na+].CC1=CC=C(S(=O)(=O)[N-]Cl)C=C1 VDQQXEISLMTGAB-UHFFFAOYSA-N 0.000 description 1
- 238000012875 competitive assay Methods 0.000 description 1
- 230000009260 cross reactivity Effects 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 230000003176 fibrotic effect Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 229940027941 immunoglobulin g Drugs 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 1
- 108010033916 pN-collagen type III Proteins 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
Abstract
It is possible by immunising animals with a peptide of the sequence I-C-E-S-C-P-T-G-G-Q-N-Y-S-P bound to an immunogenic protein to raise antibodies which can be used to determine procollagen peptide (type III) and procollagen type III selectively in body fluids.
Description
HOECHST AKTIENGESELLSCHAFT HOE 87/F 130 Dr. KH/rh Specification A method for the selective iaaunological determination of intact procollagen peptide (type III) and procollagen (type III) in body fluids, and means for carrying it out ProcolIagen pept ide (type I I I ) is the amino-terminal pro- pept ide of collagen (type III), which is cleaved off out - side the cell after secretion of the procollagen (type III) molecule. The concentration of this procollagen peptide in body fluids can be determined using a radioimmunoIogica I determination method as described in European Patent No. 4940. Knowledge of the serum concentration of the peptide allows conclusions to be drawn about the activity of fibrotic disorders, such as; for example, of the liver [Rohde, H. et al. Eur. J. Clin. Invest. £, 451 - 459 (1979)].
However, accurate selective determination of procollagen peptide (type III) in serum and other body fluids is not possible using the methods hitherto described, because the polyclonal antibodies which are used in these methods react, with different, lower affinity, with various antigens which occur in serum and some of which are breakdown products of procollagen peptide (type III) [Niemela, 0. et al. Clin.
Chim. Acta 124, 39 - 44 (1982)]. The result of this is that the serum dilution plots and the dilution plots of other body fluids are not parallel to the calibration plot constructed using pure procollagen peptide (type III), and hence it is necessary to determine the antigen content in several dilutions of each unknown sample in order to establish the antigen concentration via the 50 X intercept on the dilut ion plot.
Another disadvantage of these methods is that, because the cross-reactivity of the antibodies with rat or mouse antigen is too low, no determination of the antigen concentration in body fluids from these species has been possible.
It is possible to determine the concentrations of the antigens in rat serum using the method described by Schuppan et al. CJ. Hepatol. 3, 27-37 (1986)]. However, this method has the same disadvantage as that described for the method for human sera [Niemela et al. Clin. Chim. Acta 124, 39 44 (1982)] that the inhibition plots are not parallel for different body fluids.
This technical problem can be solved using the method of European Patent Application 0,089,008, in which the antibodies used have comparable affinities for intact procollagen peptide (type III) and its breakdown product Col 1 This method determines intact and degraded procollagen peptide (type III) together, but this results in imprecision in the diagnostic conclusions because the normal popuulation and the patient population may overlap greatly.
It has now been found, surprisingly, that when a peptide with a defined amino acid sequence is used for immunization it is possible to obtain antibodies with whose aid specific determination of intact procollagen peptide (type III) is possible. Moreover, surprisingly, using these antibodies it is possible to determine the procollagen peptide (type III) content in body fluids of rats or mice, which is of use for examining fibrosuppressive substances in animal experiments.
Thus the invention relates to a method for the immunological determination of amino-terminaI procollagen peptide (type III) using antibodies, which comprises a) immunization of animals with a peptide of the sequence I-C-E-S-C-P-T-G-G-Q-N-Y-S-P, bound to an immunogenic protein, b) obtaining from the serum the antibodies which react with intact amino-1ermina I procollagen peptide (type III) and c) determination of the amount of amino-terminaI procollagen peptide (type III) and of procollagen (type III) via the antigen-antibody complex which is formed.
The invention also relates to antibodies which are raised by immunization of animals with the abovementioned peptide which is bound to an immunogenic protein.
The invention is described in detail hereinafter, especially in its preferred embodiments. The invention is also defined in the claims.
For the preparation of the antibodies, animals, preferably rodents such as, for example, rabbits or guinea pigs, or goats and sheep are immunized with the peptide of the sequence I-C-E-S-C-P-T-G-G-Q-N-Y-S-P coupled to a suitable immunogenic protein in the presence of complete adjuvant. Rabbits are particularly preferably used. The immune response is enhanced by repeated booster injections, for example at intervals of 4 to 8 weeks. The success of immunization is checked by determining the concentration of antibodies in a radioimmunoIogica I binding assay [R. Timpl and L. Risteli, Immunochemistry of the extracellular matrix, H. Furthmayr Ed., Vol. 1, 199 (1982)3.
All immunogenic proteins are suitable as proteins to which the said peptide can be bound. Hemocyanin, albumin or polylysine are preferably used. The peptide can be prepared by methods known to the expert, as described by, - 4 for example, G. Barany and A.B. Merrifield in The Peptides Vol. 2, pp 3-254 (1980), Academic Press, or E. Brown, R.C. Sheppard and B.J. Williams, J. Chem. Soc. Perkin Transact. J./ 1 161 ( 1983).
The antibodies according to the invention can be used as serum or purified in various immunological methods, including all types of radioimmunoassay, for example sequential saturation analysis or equilibrium analysis, labeling with chloramine T or Bolton-Hunter reagents [Felber, Meth. Biochem. Anal. 22 , 1 (1974); Shelley et al., Clin. Chem. 19, 146 (1975)] as well as in other competitive binding assays such as fluorescence or enzyme immunoassays, chemiluminescence or other immunoassays. The antibodies can thus be used in immunological methods for the isolation and characterization and for the quantitative determination of procollagen peptide (type III) in tissues and body fluids. The procedure for quantitative determination makes use of methods known to the expert, by reacting a liquid sample which contains procollagen peptide (type III) with the antibodies according to the invention, and determining the amount of procollagen peptide (type III) via the antigen-antibody complex which is formed. It is of no consequence for this whether the procollagen peptide (type III) is still linked to the amino terminus of procollagen (type III) or not. The degradation products of procollagen peptide (type III), especially Col 1, which have hitherto interfered in the immunological determination, are not among the species detected by the antibodies according to the invention.
The invention is explained further in the examples which follow. Unless otherwise indicated, percentage data relate to weight.
Example 1 Preparation o'f the peptide/hemocyanin conjugate compound 100 mg of hemocyanin (dialyzed against 5 x 3 I of water and then freeze-dried) are dissolved in 3 ml of water, and 24 mg of the peptide I-C-E-S-C-P-T-G-G-Q-N-Y-S-P are added. While stirring at room temperature, a total of 1 g of Ncyclohexyl-N'-[2-(4-morpholinyl)ethyl]carbodi imide-methyl— p-1ο IuenesuIfonate is added in portions over the course of 2 hours. The reaction mixture is stirred overnight and then dialyzed against a total of 5 x 10 I of water for 24 hours, and the product is subsequently isolated by freeze-drying. 137 mg of the conjugate compound are obtained .
Example 2 Radioimmunologica I binding assay 300 ul of antiserum from rabbits immunized with the conjugate prepared as in Example 1 are, in a suitable dilution, incubated overnight with 100 μΐ of a procollagen peptide type III solution (1 ng of protein/100 μΐ, prepared as described in European Patent 4940, Example 1) which is radio125 actively labeled with I. The antigen-antibody complexes which are formed are precipitated by addition of an antiserum which is directed against immunoglobulin G of the species used for the immunization and is from a different species. Following centrifugation and decantation of the supernatant, the amount of precipitated radioactivity is determined in a γ-scinti 11 ation spectrometer.
Example 3 Radioimmunoassay 0.2 ml of the sample which is to be analyzed or of pro- 6 collagen peptide (type III) standard is incubated at 4°C overnight with an amount of antiserum (in 0.1 ml of buffer) which is limiting with respect to the amount of labeled 12 5 antigen. Addition of 0.1 ml of I-labeled procollagen peptide (type III) (contains 1 ng of protein) is followed by incubation at 4°C for 6-8 hours. The antigen-antibody complexes which are formed are precipitated by use of an antiserum directed against IgG of the species used for the immunization. Centrifugation and decantation of the supernatant are followed by determination of the precipitated radioactivity in a γ-scinti11 ation spectrometer.
It is then possible, by comparison with a calibration plot which has been constructed by using standards with different concentrations of procollagen peptide (type III), to determine the concentration of procollagen peptide (type III) in the unknown solution.
Example 4 Determination of the molecular weight distribution of the antigens which react with the antibodies according to the invention and are present in the serum of, for example, cattle and hogs ml of serum is fractionated by gel filtration chromato( R ) graphy on a Sephacryl S 300 column (1.6 x 130 cm) equilibrated in phosphate-buffered saline, PBS, containing 0.04 % Tween 20. 0.2 ml of each of the fractions (2.8 ml each) is used in the radioimmunoassay of Example 3.
Fig. 1a shows the elution profile of the antigen from porcine serum, and Fig. 1b shows that of the antigen from bovine serum, comparing with the profile of the antigen determined as in European Patent 4940. Peaks 1/1a correspond to intact procollagen type III and pN collagen type III (procollagen lacking the C-terminal end); peaks 2/2a correspond to intact amino-terminal procollagen peptide - 7 type III; peaks 3/3a correspond to Col 1 and breakdown products of amino-1ermina I procollagen peptide type III with the same molecular weight as Col 1.
Claims (10)
1. A method for the immunological determination of aminoterminal procollagen peptide (type III) using antibodies, which comprises a) immunization of animals with a peptide of the sequence > I-C-E-S-C-P-T-G-G-Q-N-Y-S-P bound to an immunogenic protein, b) obtaining from the serum the antibodies which react with intact amino-terminaI procollagen peptide (type III) and c) determination of the amount of amino-terminal procollagen peptide (type III) and of procollagen (type III) via the antigen-antibody complex which is formed.
2. The method as claimed in claim 1, wherein the peptide is bound to hemocyanin, albumin or polylysine.
3. The method as claimed in claim 1 or 2, wherein rodents or goats or sheep are immunized.
4. The method as claimed in claim 3, wherein rabbits are immunized.
5. Antibodies obtainable by immunization of animals with the peptide of the sequence I-C-E-S-C-P-T-G-G-Q-N-Y-S-P o bound to an immunogenic protein. > 6. Antibodies as claimed in claim 5, wherein the peptide is bound to hemocyanin, albumin or polylysine. Antibodies as claimed or goats or sheep are in claim 5 or 6, wherein rodents immunized. Antibodies as claimed in claim 7, wherein rabbits are i mmun i zed. A compound composed of the peptide of the sequence I-C-E-S-C-P-T-G-G-Q-N-Y-S-P bound to an immunogenic protein.
6. 10. The use of the compound claimed in claim 9 as an antigen.
7. 11. A method according to Claim 1 for the immunological determination of amino-terminal procollagen peptide (type 111) using antibodies, substantially as hereinbefore described and exemplified. j
8. 12. An antibody according to Claim 5, substantially as ) hereinbefore described and exemplified.
9. 13. A compound according to Claim 9, substantially as hereinbefore described and exemplified.
10. 14. Use according to Claim 10, substantially as hereinbefore described and exemplified.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19873714634 DE3714634A1 (en) | 1987-05-02 | 1987-05-02 | METHOD FOR SELECTIVE IMMUNOLOGICAL DETERMINATION OF INTACT PROCOLLAGEN PEPTIDE (TYPE III) AND PROCOLLAGEN (TYPE III) IN BODY LIQUIDS AND MEANS TO IMPLEMENT IT |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| IE881289L IE881289L (en) | 1988-11-02 |
| IE63385B1 true IE63385B1 (en) | 1995-04-19 |
Family
ID=6326686
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| IE128988A IE63385B1 (en) | 1987-05-02 | 1988-04-29 | A method for the selective immunological determination of intact procollagen peptide (Type III) and procollagen (Type III) in body fluids and means for carrying it out |
Country Status (10)
| Country | Link |
|---|---|
| EP (1) | EP0298210B1 (en) |
| JP (1) | JPH0780913B2 (en) |
| AT (1) | ATE98377T1 (en) |
| DE (2) | DE3714634A1 (en) |
| DK (1) | DK168872B1 (en) |
| ES (1) | ES2061545T3 (en) |
| FI (1) | FI95579C (en) |
| IE (1) | IE63385B1 (en) |
| NO (1) | NO175639C (en) |
| PT (1) | PT87375B (en) |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6010862A (en) * | 1987-11-06 | 2000-01-04 | Washington Research Foundation | Methods of detecting collagen type III degradation in vivo |
| DE4225038C2 (en) * | 1992-07-29 | 1995-11-30 | Boehringer Mannheim Gmbh | Production and use of antibodies against collagen |
| DK0711415T3 (en) * | 1993-07-28 | 1999-04-26 | Roche Diagnostics Gmbh | Immunoassay for detection of collagen or collagen fragments |
| GB9506050D0 (en) | 1995-03-24 | 1995-05-10 | Osteometer A S | Assaying collagen fragments in body fluids |
| DE69520111T2 (en) | 1994-10-17 | 2001-09-06 | Osteometer Biotech As Herlev | Estimation of the fragmentation pattern of collagen in body fluids and diagnosis of disorders related to collagen metabolism |
| US6107047A (en) * | 1996-03-21 | 2000-08-22 | Osteometer Biotech A/S | Assaying protein fragments in body fluids |
| GB9617616D0 (en) | 1996-08-22 | 1996-10-02 | Osteometer Biotech As | Assaying protein fragments in body fluids |
| DE69705423T2 (en) | 1996-12-09 | 2002-05-16 | Osteometer Biotech As Herlev | SANDWEIGHT TEST FOR DETECTING COLLAGEN FRAGMENTS |
| US6117646A (en) * | 1997-09-22 | 2000-09-12 | Osteometer Biotech A/S | Assaying protein fragments in body fluids |
| ATE260298T1 (en) | 1998-05-28 | 2004-03-15 | Bayer Healthcare Ag | MONOCLONAL ANTIBODIES AND ASSAY FOR DETERMINATION OF N-TERMINAL PROCOLLAGEN PROPEPTIDE TYPE III (PIIINP) |
| US6602980B1 (en) | 1998-06-19 | 2003-08-05 | Washington Research Foundation | Collagen type III synthetic peptides for collagen resorption assays |
| US6916903B2 (en) | 1998-06-19 | 2005-07-12 | Washington Research Foundation | Collagen type III synthetic peptides for collagen resorption assays |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE2816841A1 (en) * | 1978-04-18 | 1979-10-31 | Max Planck Gesellschaft | RADIOIMMUNOLOGICAL DETERMINATION OF PROCOLLAGEN (TYPE III) AND PROCOLLAGEN PEPTIDE (TYPE III) |
| DE3209149A1 (en) * | 1982-03-13 | 1983-10-06 | Hoechst Ag | METHOD FOR THE COMMON IMMUNOLOGICAL DETERMINATION OF PROCOLLAGEN PEPTIDE (TYPE III) AND PROCOLLAGEN PEPTIDE COL 1 (TYPE III) AND METHOD FOR THE PRODUCTION OF ANTI-PROCOLLAGEN PEPTIDE COL 1 (TYPE III) SERUM |
-
1987
- 1987-05-02 DE DE19873714634 patent/DE3714634A1/en not_active Withdrawn
-
1988
- 1988-04-27 ES ES88106747T patent/ES2061545T3/en not_active Expired - Lifetime
- 1988-04-27 DE DE88106747T patent/DE3886109D1/en not_active Expired - Fee Related
- 1988-04-27 EP EP88106747A patent/EP0298210B1/en not_active Expired - Lifetime
- 1988-04-27 AT AT88106747T patent/ATE98377T1/en not_active IP Right Cessation
- 1988-04-28 JP JP63104474A patent/JPH0780913B2/en not_active Expired - Lifetime
- 1988-04-28 DK DK232988A patent/DK168872B1/en not_active IP Right Cessation
- 1988-04-29 NO NO881904A patent/NO175639C/en not_active IP Right Cessation
- 1988-04-29 PT PT87375A patent/PT87375B/en active IP Right Grant
- 1988-04-29 IE IE128988A patent/IE63385B1/en not_active IP Right Cessation
- 1988-04-29 FI FI882018A patent/FI95579C/en not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| FI882018A (en) | 1988-11-03 |
| JPH0780913B2 (en) | 1995-08-30 |
| IE881289L (en) | 1988-11-02 |
| ATE98377T1 (en) | 1993-12-15 |
| ES2061545T3 (en) | 1994-12-16 |
| DE3886109D1 (en) | 1994-01-20 |
| NO881904L (en) | 1988-11-03 |
| PT87375B (en) | 1992-08-31 |
| NO175639B (en) | 1994-08-01 |
| DK168872B1 (en) | 1994-06-27 |
| EP0298210B1 (en) | 1993-12-08 |
| DE3714634A1 (en) | 1988-11-17 |
| JPS63292061A (en) | 1988-11-29 |
| NO881904D0 (en) | 1988-04-29 |
| DK232988D0 (en) | 1988-04-28 |
| PT87375A (en) | 1989-05-31 |
| FI95579B (en) | 1995-11-15 |
| FI882018A0 (en) | 1988-04-29 |
| FI95579C (en) | 1996-02-26 |
| NO175639C (en) | 1994-11-09 |
| DK232988A (en) | 1988-11-03 |
| EP0298210A3 (en) | 1991-05-15 |
| EP0298210A2 (en) | 1989-01-11 |
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