Generation of monoclonal anti-dermatan sulphate antibodies cross-reacting to calf thymus DNA.

ABSTRACT

Monoclonal anti-dermatan sulphate antibodies were produced with hybridization after immunizing BALB/c mice with dermatan sulphate (DS). These antibodies were tested for cross-reactivity to double-stranded DNA (dsDNA) and heat-denatured DNA (ssDNA). Five clones had binding activity to DS, dsDNA and ssDNA in enzyme-linked immunosorbent assay, but no clones binding to DS alone were produced. Two (2H8 and 3C4) of the five clones were selected for analysis. Their binding activity to DS, ssDNA and dsDNA were inhibited by DS, dsDNA and ssDNA. DNase I treatment abolished the binding activity to dsDNA and ssDNA, completely, but had no effect on the binding activity to DS. On the other hand, chondroitinase ABC treatment of the solid-phased DS augmented the binding to DS. The inhibition assay using digested fractions suggested that the epitope recognized by 2H8 and 3C4 clones could not be in delta Di-4S or delta Di-diSB but in the linkage regions of DS.