Generation and identification of natural monoclonal antibodies against low-density lipoprotein.

摘要

ABSTRACT Low-density lipoprotein (LDL) is a major trigger of atherosclerotic lesions. Here we report the production of natural mouse IgM monoclonal antibodies (MAbs) against LDL with high specificity and activity, which could provide a potential value for research on lipid metabolism and atherosclerosis progression. BALB/c mice were raised in specific pathogen-free conditions and fed a high cholesterin diet; the splenocytes from these mice were directly fused with Sp2/0 myeloma cells using standard hybridoma production techniques. Resulting hybridomas producing anti-LDL antibodies were screened by enzyme-linked immunosorbent assay (ELISA) and isotyped. As a result, two hybridoma cell lines, named 5G8 and 2H7, were developed, which could secrete anti-LDL MAbs stably. Both of them belonged to the IgM subclass. The specificity of the MAb was determined based on its activity in Western blot and immunoprecipitation analysis.