Generation and identification of natural monoclonal antibodies against low-density lipoprotein.
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.
Feng
Xuyang
X
Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi Province, China.
Xu
Ruifen
R
Gao
Yan
Y
He
Zheng
Z
Li
Chengxiang
C
Wang
Haichang
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D000918:Antibody Specificity; D002791:Cholesterol, Dietary / Q000008:administration & dosage; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology / Q000302:isolation & purification*; D008077:Lipoproteins, LDL / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D002791:Cholesterol, Dietary; D007075:Immunoglobulin M; D008077:Lipoproteins, LDL