Monoclonal Antibodies to the Large Chondroitin Sulphate Proteoglycan from Bovine Temporomandibular Joint Disc

Abstract

Four hybrid cell lines producing monoclonal antibodies (designated AC2, AH12, DB10 and DD11) were derived from mice immunized with the large chondroitin sulphate proteoglycan isolated and purified from the bovine temporomandibular joint disc. The epitopes were partially characterized by enzyme-linked immunosorbent assays and staining patterns on immunoblots of intact proteoglycans and digests made with glycosidases and proteinases. All four monoclonal antibodies appeared to recognize some form of keratan sulphate although the epitopes for two (AC2 and DD11) were probably identical. One antibody (AH12) showed almost no reactivity with corneal keratan sulphate but stained a small keratan sulphate proteoglycan extracted from the disc, in addition to the large chondroitin sulphate proteoglycan. These antibodies were used for immunohistochemical staining of sections of the disc and showed that keratan sulphate associated with the large chondroitin sulphate proteoglycan was concentrated inside and away from the periphery of the structure but close to the inferior and superior surfaces, in a pattern which may reflect the adaptation of the extracellular matrix to the mechanical stresses placed on it by mastication.