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A FIBRIN-SPECIFIC MONOCLONAL ANTIBODY WITH EPITOPE LOCALIZED IN FIBRIN FRAGMENT BBETA118-134: ITS APPLICATION TO QUANTIFICATION OF SOLUBLE FIBRIN
Abstract number: P-W-382
Lugovskoy1 E.V., Kolesnikova1 I.N., Gritsenko1 P.G., Lugovskaya1 N.E., Komisarenko1 S.V.
1Department of Molecular Immunology, Palladin Institute of Biochemistry, Kiev, Ukraine
How-to-cite Lugovskoy EV, Kolesnikova IN, Gritsenko PG, Lugovskaya NE, Komisarenko SV. A FIBRIN-SPECIFIC MONOCLONAL ANTIBODY WITH EPITOPE LOCALIZED IN FIBRIN FRAGMENT BBETA118-134: ITS APPLICATION TO QUANTIFICATION OF SOLUBLE FIBRIN. J Thromb Haemost 2007; 5 Supplement 2: P-W-382
Abstract
Introduction: Upon thrombin-catalyzed cleavage of fibrinogen in blood plasma the soluble fibrin is formed. The latter serves as a molecular marker of activation of blood coagulation system and of the risk of thrombus formation.
Methods: Standard hybridoma technique was used for production of monoclonal antibodies (mAbs), which were isolated from the cultural media by affinity chromatography on protein G- and fibrin- Sepharoses.
Results: Using partly denaturated human fibrin we obtained mAb 1A-5C reacting with fibrin desAA, fibrin desAABB and not reacting with fibrinogen, X-fragment, D-dimer and t-NDSK. Immunoblot analysis showed that mAb 1A-5C reacted with fibrin desAABB Bbeta-chain 15-461 and did not react with Bbeta 15-118 of t-NDSK and Bbeta 134-461 of D-dimer. These data testify that epitope for mAb 1A-5C is localized at fibrin region Bbeta118-134. Thus this region is the neoantigenic determinant being exposed during fibrinogen-fibrin transformation. We have elaborated double-sandwich ELISA for quantification of soluble fibrin in human blood plasma using fibrin-specific mAb 1A-5C as a "catch" and our mAb II-4d as a "tag" [Lugovskoy E.V., et al. Thromb. Res. 2004. 113, N 3-4, P.251-259]. The method has been used for soluble fibrin quantification in blood of pregnant women with the risk of fetal loss.
Conclusions: Fibrin-specific mAb 1A-5C has been obtained with epitope being localized in human fibrin fragment Bbeta118-134. This mAb was used in double-sandwich ELISA as a "catch"-antibody for soluble fibrin quantification in human blood plasma.
To cite this abstract use the following format:
Journal of Thrombosis and Haemostasis 2007; Volume 5, Supplement 2: abstract number
Session Details
| Date: |
01/08/2007
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| Time: |
00:00-00:00
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| Session name: |
XXIst ISTH Congress |
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| Location: |
Oxford, UK |
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