DEVELOPMENT OF MURINE MONOCLONAL ANTIBODIES TO PNEUMOCOCCAL POLYSACCHARIDE ANTIGEN

Abstract

Streptococcus pneumoniae is a major cause of mortality in infants, children and the elderly. Despite the availability of excellent antimicrobial therapy and adequate care systems, respiratory diseases and invasive infections caused by pneumococci still comprise a major health problem. Thus to overcome this, murine monoclonal antibodies were developed using pneumococcal polysaccharide as the antigen. Polyclonal MAb was developed against different serotypes namely 4, 6B, 9V, 14, 18C, 19F, 23F. Hybridoma cell line developed against Serotype 4 showed maximum fluorescence absorbance units when detected, using Rapid multiplex assay. Single cell cloning against Serotype 4 hybridoma cell line was done by Limiting dilution. A stable single hybridoma cell line was developed producing Monoclonal antibody against pneumococcal polysaccharide Type 4 serotype. The expression of the murine monoclonal antibody was assayed using SDS- PAGE (Reducing). Presence of heavy chain at 50kDa and light chain at 25kDa in reducing condition confirmed presence of MAb.