Document Detail

Calcium ion-dependent monoclonal antibody against human fibrinogen: preparation, characterization, and application to fibrinogen purification.
MedLine Citation:
PMID:  7495075     Owner:  NLM     Status:  MEDLINE    
We have produced a high-affinity monoclonal antibody classified as IgG1 with kappa-type light chains that recognizes the calcium ion(Ca2+)-dependent conformation of the D-domain of human fibrinogen. Binding of fibrinogen in solution to the insolubilized antibody increased in the presence of increasing concentrations of up to 2 mM Ca2+, the half-maximal binding being reached at 130 microM Ca2+. The dissociation constant was estimated to be 1.6 x 10(-8) M at 2 mM Ca2+. The antibody was found also to be dependent on other divalent metal ions including Zn2+, Mn2+, Co2+ and Cu2+, but not Ba2+, Mg2+ or Sr2+. The synthetic Gly-Pro-Arg-Pro-amide peptide, which has recently been shown to bind to close proximity to the calcium binding site in the D-domain, was unable to elicit the conformation for the antigen to be recognized by this antibody. This antibody was found to be a suitable ligand for the immunoaffinity chromatography of normal and abnormal fibrinogens directly from citrated plasma depleted of the vitamin K-dependent proteins or heparinized plasma by eliminating the precipitation procedure widely adopted in conventional techniques of fibrinogen purification. Indeed, fibrinogen Marburg I with the A alpha chains depleted of the carboxy-terminal A alpha(461-610) residue segment has been purified by this technique, although this dysfibrinogen was difficult to purify by conventional precipitation techniques.
M Takebe; G Soe; I Kohno; T Sugo; M Matsuda
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Thrombosis and haemostasis     Volume:  73     ISSN:  0340-6245     ISO Abbreviation:  Thromb. Haemost.     Publication Date:  1995 Apr 
Date Detail:
Created Date:  1996-01-11     Completed Date:  1996-01-11     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7608063     Medline TA:  Thromb Haemost     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  662-7     Citation Subset:  IM    
Division of Hemostasis and Thrombosis Research, Jichi Medical School, Tochigi, Japan.
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MeSH Terms
Antibodies, Monoclonal / immunology*,  isolation & purification
Calcium / chemistry*,  immunology
Epitopes / chemistry,  immunology
Fibrinogen / chemistry,  immunology*,  isolation & purification
Immunosorbent Techniques
Protein Conformation
Sensitivity and Specificity
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Epitopes; 7440-70-2/Calcium; 9001-32-5/Fibrinogen

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