PRODUCTION AND SPECIFICITY OF ANTIBODIES AGAINST AMINOTERMINAL REGION IN TYPE-3 COLLAGEN

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  • 1 January 1976
    • journal article
    • research article
    • Vol. 31  (1) , 57-65
Abstract
A cross-linked fragment (peptide T1X) with a MW of 13,000 could be isolated from a tryptic digest of insoluble type III collagen of calf skin. Peptide T1X was conjugated on to bovine serum albumin by glutaraldehyde and used for immunization of rabbits. The antisera reacted in passive hemagglutination and radioimmune assay with peptide T1X, type III collagen and its constituent .alpha.1(III) chain. Little or no reaction was observed with type I collagen and .alpha.1(I) chain. While rabbit antisera to neutral salt-soluble type III collagen also showed a strong binding for 125I-labeled peptide T1X, much less reaction was observed with antisera to type I collagen. The antigenicity of type III collagen was largely destroyed by pepsin treatment, suggesting that it resided in non-helical segments. A fragment of peptide T1X produced by digestion with collagenase retained antigenic activity. The amino-terminal region of type III collagen apparently contains strong antigenic determinants located in a non-helical sequence of about 16 amino acids. Antibodies to these antigenic determinants were purified and rendered specific for type III collagen by immunoadsorption. The antibodies stained in indirect immunofluorescence tests particularly those regions in various connective tissues which are rich in reticulin fibers. Different staining patterns were observed with antibodies to type I collagen.