Production and specificity of antibodies against the aminoterminal region in type III collagen.
A cross-linked fragment (peptide T1X) with a molecular weight 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 haemagglutination and radioimmune assay with peptide T1X, type III collagen and its constituent alpha1(III) chain. Little or no reaction was observed with type I collagen and alpha1(I) chain. While rabbit antisera to neutral salt-soluble type III Collagen also showed a strong binding for 125I-labelled 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 data indicated that the aminoterminal region of type III collagen contains strong antigenic determinants located in a non-helical sequence of about sixteen 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 fibres. Different staining patterns were observed with antibodies to type I collagen.
D000906:Antibodies / Q000302:isolation & purification; D000917:Antibody Formation; D000918:Antibody Specificity; D003094:Collagen / Q000276:immunology*; D003429:Cross Reactions; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D007163:Immunosorbent Techniques; D010455:Peptides / Q000276:immunology*
D000906:Antibodies; D000939:Epitopes; D010455:Peptides; D003094:Collagen