Preparation of monoclonal antibodies against gamma-type phospholipase A2 inhibitors and immunodetection of these proteins in snake blood
Li, JingjingXiong, YingSun, ShiminYu, LehanHuang, Chunhong
Abstract Background: The gamma-type phospholipase A2 inhibitor (LI) is a natural protein commonly found in snake serum, which can neutralize pathophysiological effects of snake venom phospholipases A2. Therefore, this protein is a potential candidate to the development of a novel antivenom. To the best of our knowledge, there is no antibody currently available for PLI identification and characterization. Methods: Bioinformatics prediction of epitope using DNAStar software was performed based on the sequence of Sinonatrix annularis PLI (SaPLI). The best epitope 151CPVLRLSNRTHEANRNDLIKVA172 was chosen and synthesized, and then conjugated to keyhole limpet hemocyanin and bovine serum albumin for use as an immunogen and plate-coating antigen, respectively. Results: Eighteen IgG anti-PLI mAb hybridoma cell strains were obtained, and all the mAbs had positive interaction with recombinant His6-PLI and natural SaPLI. Moreover, the mAb from 10E9 strain was also successfully used for the immunodetection of other snake serum PLIs. cDNA sequence alignment of those PLIs from different snake species showed that their epitope segments were highly homologous. Conclusions: The successful preparation of anti-PLImAb is significant for further investigation on the relationship between the structure and function of PLIs, as well as the interaction between PLIs and PLA2s.
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