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《Chinese Journal of Biochemical Pharmaceutics》 2010-01
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Preparation and identification of anti-HIV-1 gp120 monoclonal antibody

ZHANG Yong-chen,WEN Jian,ZHANG Liang,ZHAO Lei(The N0.2 Hospital Affiliated to Southeast University,Nanjing 210003,China)  
Purpose To Prepare anti-HIV-1 gp120 monoclonal antibodies and to identify the specificity of antibodies in order to provide technique for preparing HIV remedial antibodies.Methods The gene fragment of HIV-1 gp120 was connected to PEGX-4T-2 prokaryotic expressing vector.The vector was cut by enzyme.GST-HIV protein was expressed by E.coli XL1-blue.The BALB/c mice were immunized with purified GST-HIV antigen,and then the fusion of mice spleen cell and myeloma cell SP2/0 was executed as the routine cell-fusion technique.Positive cells were screened by Indirect ELISA.Immuno-blotting assay and Western blot identified the specificity of antibodies.Results External gene section from the recombinant plasmid by sequencing showed the same size of HIV-1 gp120 gene sequences.An external expressed protein band of 32 KD was obtained after purified protein SDS-PAGE electrophoresis.It indicated that six strains of hybridoma cells secreting special monoclone antibodies had been obtained.ELISA results showed that six strains monoclonal antibodies only reacted to HIV-1 gp120 antigen.Western blot results showed that a band with molecular weight 32 kDa was obtained,which could interact with HIV-1 gp120 monoclonal antibody.Conclusion Six strains of hybridoma cells secreting special monoclone antibodies had been obtained.The prepared monoclonal antibodies have established a basis for HIV remedial antibody.
【Fund】: 江苏省自然科学基金(BK2004013);; 南京市社会发展项目(200601046)
【CateGory Index】: R392;R512.91
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