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Abstract
Extracts of normal and keratoconus human corneas were assayed for keratan sulfate (KS) and for the protein core of corneal keratan sulfate proteoglycan (KSPG), using a solid-phase immunoassay. Aliquots from guanidine-HCl extracts of four to six pooled corneas were adsorbed to nitrocellulose membrane and assayed for binding of a monoclonal antibody against KS or of an affinity-purified rabbit polyclonal antibody against the KSPG protein core using 125I-labeled secondary antibodies. The amount of antigen in the extracts was estimated from standard curves with purified bovine KSPG. Extracts from keratoconus corneas contained only an average of 48% as much (P less than 0.001) KS-antigen as normal corneal extracts when assayed with anti-KS monoclonal antibody. There was no significant difference in the amount of KSPG core protein antigens in keratoconus and normal corneal extracts. These results suggest that corneas with keratoconus contain a form of KSPG that contains fewer keratan sulfate chains or in which the keratan sulfate has a modified structure.