[Expression of human ScFv against human immunodeficiency virus type 1 gp120].
In order to generate human antibody against HIV-1, using synthetic peptide from HIV-1 gp120 v3 loop as antigen, we screened the recombinant phage antibodies by immunoaffinity and selected positive clones carrying anti-gp120 ScFv gene. The phage DNA was extracted and transformed to E. coli HB2151 that allowed to express soluble ScFv proteins by inducing of IPTG. The results of SDS-PAGE and Western blot showed that the molecular weight of the expressed product is about 28 KD and exhibited anti-c-myc activity. ELISA and Dot blot analysis displayed that the soluble ScFv specifically bound to gp120 and had a higher specificity and affinity. Competitive ELISA further demonstrated that the product was specific to gp120.
He
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Department of Virology, 302 Hospital of PLA, Beijing 100039.
Mao
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Zhu
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D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis* / Q000235:genetics; D015497:HIV-1 / Q000235:genetics; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D057127:Single-Chain Antibodies
D000914:Antibodies, Viral; D004274:DNA, Recombinant; D015699:HIV Envelope Protein gp120; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C114805:scFv fragment SW1