Preparation of antibodies with glycosidase activity using in vitro and in vivo immunizations

Abstract

Catalytic antibodies were prepared using a transition state analog and monoclonal antibody technology. A synthetic hapten, mimicking transient state of a glycosidic bond, was synthesized. Three approaches, in vitro immunization, SJL/J mice immunization and conventional BALB/c mice immunization, were employed to prepare hybridoma clones producing antibodies with catalytic activities. All three approaches produced a significant number of hybridoma clones producing hapten binding antibodies. The catalytic activities of the antibodies were examined using hybridoma culture supernatants and p-nitro-alpha-D-glucopyranoside. The fractions of the hybridoma clones producing antibodies with the glycosidase activity, however, were significantly different among the methods employed. Most of the hybridoma clones obtained from in vitro and SJL/J mice immunizations happened to produce antibodies with glycosidase activity, while most of those hom conventional BALB/c mice immunization did not.