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A preliminary study on single chain fragments variable (ScFv) antibody against HIV-1 integrase by phage surface displaying

Jianhui Zhang, Meng Li, Baoguang Han, Xiaoguo Song, Yuying Feng, Shigan Ling, Xiankai Ma

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Abstract

A combinatory single-chain fragment variable (ScFv) cDNA library was constructed from the splenic mRNA of a mouse immunized with IN protein. The library cDNA was cloned into a phagemid vector pCANTAB5E and introduced into E.coli TG1, 3.5×10~(5) ampicillin resistant clones were obtained. After four rounds of panning against IN coated on immune plate, 76 IN-binding phage-ScFvs were selected. DNA Sequence analyzed from a randomly selected clone proved to be in accordance with mouse antibody. Phage-ScFvs against HIV-1 IN were collected.

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What this paper is about

A combinatory single-chain fragment variable (ScFv) cDNA library was constructed from the splenic mRNA of a mouse immunized with IN protein. The library cDNA was cloned into a phagemid vector pCANTAB5E and introduced into E.coli TG1, 3.5×10~(5) ampicillin resistant clones were obtained. After four rounds of panning against IN coated on immune plate, 76 IN-binding phage-ScFvs were selected. DNA Sequence analyzed from a randomly selected clone proved to be in accordance with mouse antibody. Phage-ScFvs against HIV-1 IN were collected.

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Available abstract

A combinatory single-chain fragment variable (ScFv) cDNA library was constructed from the splenic mRNA of a mouse immunized with IN protein. The library cDNA was cloned into a phagemid vector pCANTAB5E and introduced into E.coli TG1, 3.5×10~(5) ampicillin resistant clones were obtained. After four rounds of panning against IN coated on immune plate, 76 IN-binding phage-ScFvs were selected. DNA Sequence analyzed from a randomly selected clone proved to be in accordance with mouse antibody. Phage-ScFvs against HIV-1 IN were collected.

Key concepts: Integrase, Phagemid, Phage display, Panning (audio), Virology, Complementary DNA, Molecular biology, Biology

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