Immunochemical studies of early alpha chain crosslinking in human fibrin

Thesis/Dissertation · Thu Jan 01 04:00:00 EST 1987

OSTI ID:7027153

Sobel, J H

Immunochemical studies were conducted to characterize the earliest covalent ..cap alpha.. chain associations catalyzed by Factor XIII/sub a/ during in vitro fibrin formation. Preparations of purified fibrinogen and Factor XIII were incubated under crosslinking conditions for increasing periods of time and the resulting fibrins subjected to immunoblotting using the characterized monoclonal antibodies, F-103 (anti-A..cap alpha.. number259-276) and F-102 /anti-A..cap alpha.. number540-554). Data obtained for the incorporation of monomeric ..cap alpha.. chains into higher molecular weight immunoreactive species indicate that ..cap alpha.. chain crosslinking begins within the first ten minutes of clotting. Early crosslinked ..cap alpha.. chain species that formed within thirty minutes of in vitro crosslinking were isolated from preparations of reduced, /sup 3/H-S-carboxymethylated fibrin by Sepharose CL-4B gel filtration. Cyanogen bromide treatment of this material resulted in the release of crosslinked and non-crosslinked fragments which could be separated from one another by Sephadex G-150 gel filtration. Derivatives of interest were identified in the column effluent by immunoblotting with F-103 and F-102 and by radioimmunoassays in which polyclonal antisera that detect the known (A) ..cap alpha.. chain crosslinking regions, number241-476 (CNRr VIII) and number518-584 (CNBr X), were employed.

• Research Organization: Columbia Univ., New York (USA)
• OSTI ID: 7027153
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
ANTIBODIES
CHEMICAL REACTIONS
COAGULANTS
CROSS-LINKING
DRUGS
FIBRIN
FILTRATION
HEMATOLOGIC AGENTS
HEMOSTATICS
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MAN
MOLECULAR WEIGHT
MONOCLONAL ANTIBODIES
ORGANIC COMPOUNDS
POLYMERIZATION
PRIMATES
PROTEINS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNOLOGY
SCLEROPROTEINS
SEPARATION PROCESSES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES