Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antibodies, Viral / biosynthesis*
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Antibodies, Viral / genetics
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Cloning, Molecular
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DNA, Recombinant / genetics
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Escherichia coli / genetics
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Genetic Vectors
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HIV Envelope Protein gp120 / biosynthesis*
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HIV Envelope Protein gp120 / genetics
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HIV-1 / genetics
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Immunoglobulin Fragments / genetics
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Immunoglobulin Fragments / immunology*
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Immunoglobulin Variable Region / genetics
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Immunoglobulin Variable Region / immunology*
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Polymerase Chain Reaction
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / genetics
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Single-Chain Antibodies
Substances
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Antibodies, Viral
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DNA, Recombinant
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HIV Envelope Protein gp120
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Immunoglobulin Fragments
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Immunoglobulin Variable Region
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Recombinant Proteins
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Single-Chain Antibodies
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scFv fragment SW1