PMID: 6171570Jan 10, 1982Paper

Antibodies against the predominant glycosaminoglycan of the mammalian cornea, keratan sulfate-I.

The Journal of Biological Chemistry
G W ConradM L Woo

Abstract

Antibodies have been made in rabbits against bovine corneal keratan sulfate proteoglycan. Antisera were titered by their ability to agglutinate sheep red blood cells that had been coated with the proteoglycan. Immune antisera, but not preimmune sera, agglutinate coated cells. Uncoated cells are not agglutinated by either serum. Immune agglutination is inhibited by prior incubation of antiserum with the intact corneal proteoglycan fraction or with 2-mercaptoethanol. Immune agglutination is also sharply reduced by the glycosaminoglycans, keratan sulfate-I (corneal type), and keratan sulfate-II (cartilage type). Desulfated keratan sulfate-I is somewhat less effective as an inhibitor than keratan sulfate-I. In contrast, chondroitin 4- and 6-sulfates, heparin, and hyaluronic acid do not interfere with immune agglutination. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by electroblot transfer of the proteins to nitrocellulose paper, incubation with antisera, and reaction with 125I-protein A suggest that the proteoglycan fraction contains high molecular weight antigenic components (Mr = approximately 300,000) whose mobility is sharply decreased by incubation with keratanase to that corresponding to mo...Continue Reading

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