HIGH-PRESSURE-INDUCED MODULATION OF THE ANTIGENIC INTERACTIONS BETWEEN 2 BETA-GALACTOSIDASES AND ANTI-BETA-GALACTOSIDASE ANTIBODIES

Disruption of antigen-antibody complexes often requires drastic condit ions. High-pressure application to such complexes would lead to the ru pture of antigen-antibody interactions as suggested by Olson et al. Th e results presented here attempted to determine the usuability of high pressure as an alternative antigen-antibody separation ration process using a beta-galactosidase-anti-beta-galactosidases complex as a mode l. For this purpose, pressure- changes concerning the biologic functio ns of two beta-galactosinases and polyclonal-related antibodies were s tudied in the 0-500 MPa pressure range for temperatures varying from 4 to 35 degrees C. After pressure release, catalytic activity of the be ta-galactosidases was measured. In addition, enzyme-linked immunosorbe nt assay (ELISA) was used to examine antibody recognition after these proteins had been pressure-treated. Recognition of antigens by the pre ssurized antibodies was also investigated by ELISA. This work allowed us to show the possible design of a high-pressure separation process w ith a beta-galactosidase-anti-beta-galactosidase tandem (or fusion pro teins possessing Escherichia coli beta-galactosidase as a tag). For pr essures below 250 MPa within the temperature range investigated, no ir reversible drastic loss of activity or dramatic alterations in antigen -antibody recognition was observed. Moreover; interesting pressure and temperature effects leading to changes in antigenic reactivity were o bserved and are discussed.