Production and partial characterization of monoclonal antibodies to the neotype strain of Mycobacterium bovis

Abstract

SUMMARY Six monoclonal antibodies (mab) to virulent Mycobacterium bovis ATCC 19210 were produced, using a suspension of heat-inactivated whole cells. Immunoglobulin isotype for mab VMB6, VMB73, and VMB93 was IgG1, and for VMB31, VMB99, and VMB119, it was IgG2a. Monoclonal antibodies were examined for cross-reactivity to M tuberculosis, M kansasii, M fortuitum, M paratuberculosis, M avium serovars 1, 2, 4, 8, and 10, M chelonei, M phlei, M scrofulaceum, M smegmatis, Nocardia asteroides, and Rhodococcus equi. Monoclonal antibodies could be grouped on the basis of binding activity by elisa and immunoblot analysis, in which mab VMB6, VMB31, and VMB119 had binding activity to M bovis; mab VMB93 and VMB99 detected M bovis and M tuberculosis antigens, and mab VMB73 reacted with other mycobacterial species, as well as with N asteroides and R equi. Apparent molecular mass of antigens was 30 to 25 kilodaltons (kD) for VMB6, VMB31, and VMB119 and 63 kD for VMB93 and VMB99, and ranged from > 200 to 31 kD for VMB73, as estimated by immunoblot analysis. Monoclonal antibody binding activity to 18 field isolates of M bovis was evaluated, using elisa. Each of 18 field isolates was detected, using mab VMB6, VMB31, or VMB119; 10 isolates were detected, using mab VMB93/VMB99, and 14 were detected by use of mab VMB73. Use of mab in elisa failed to detect antigens from M bovis strain AN-5.