ABSTRACT
Human Type I collagen can be extracted from spleen with acetic acid or from skin by incubation with pepsin; these extracts are then purified by amino acid analysis and gel electrophoresis. Such antibodies can be utilized to quantify human type I collagen in an enzyme-linked immunoassay (ELISA). Measurement in all ELISA tests depends on the presence of an active enzyme bound to one of the components of the antigen antibody reaction. The inhibition type of the ELISA test is relatively unaffected by minor contaminants in either antigen or antibody. Since contaminants are a major problem in both collagen preparations and anticollagen antibodies, the inhibition test is best suited to measure Type I collagen. The classical approaches for the quantitation of collagen take advantage of unique features of collagen such as the presence of hydroxyproline1or the sensitivity of the protein to bacterial collagenase.
