Monoclonal Antibody Produced Against Calf Thymus Histone

Abstract

The localization and modification of histone play a prime role in the regulation of unelucidated biochemical pathways, such as apoptosis and cell proliferatory-inhibitory activities. Thus studying histone reveals cell proliferation and its biochemical processes. Therefore anti-histone antibody production is important in order to isolate, purify, identify, and separate histone from cultured cells. In order to produce anti-histone antibodies by hybridoma, 8-week-old female BALB/c mice were immunized with commercially procured whole calf thymus histone. Subsequently the spleen cells of the immunized BALB/c and SP2/O-AG-14 myeloma cells were fused using polyethylene glycol, which resulted in 16 clones. Out of these 16 clones, only one, 1D7, was found to be specific to our antigen (calf thymus histone) by ELISA. The supernatant of the positive clone 1D7 was used in immunoblotting to characterize the specificity of the produced monoclonal antibody. Immunoblotting confirmed that the monoclonal antibody produced from 1D7 clone is specific to histone H1 epitope present in the calf thymus. Our results suggest that to generate more potent monoclonal antibody reagent specific for many different epitopes present on histone, a larger number of fusions are required. Further, if such potent monoclonal antibody reagent is produced, it will be an indispensable tool in elucidating the role of H1 in apoptosis and cell proliferatory-inhibitory activities. This reagent can be used for diagnosis of antihistone antibody in SLE patient's serum, for testing the viability of cultured monocytes, and for localization of histone in metaphase chromosomes.