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2006•Jiangsu Medical JournalRequires access

Construction of a single chain Fv phage display library against human B-lymphoma

Xiaochun Yang

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Abstract

Objective To construct a single chain Fv(ScFv) phage display library against(human) B-lymphoma.Methods The mRNA was extracted from the spleen cells of the immunized(BALB/c) mice with Raji B-lymphoma cell strain and cDNA was synthesized by reverse transcriptase.The heavychain and light-chain variable region genes(V_H and V_L)of immunoglobulin were amplified by PCR and joined by a DNA linker to encode peptide as a single chain Fv(ScFv) fragment.ScFv fragments were amplified and cloned into the phagemid vector pHEN1 and the phage display library was constructed.Results The ScFv phage display library had a capacity of approximately 3.4×10~6,with 100% of the phagemids containing ScFv gene insertion as demonstrated by PCR.Conclusion The ScFv phage display library against human B-lymphoma was successfully constructed.The present study can be used for succeeding screening of specific antibody against human B-lymphoma.

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What this paper is about

Objective To construct a single chain Fv(ScFv) phage display library against(human) B-lymphoma.Methods The mRNA was extracted from the spleen cells of the immunized(BALB/c) mice with Raji B-lymphoma cell strain and cDNA was synthesized by reverse transcriptase.The heavychain and light-chain variable region genes(V_H and V_L)of immunoglobulin were amplified by PCR and joined by a DNA linker to encode peptide as a single chain Fv(ScFv) fragment.ScFv fragments were amplified and cloned into the phagemid vector pHEN1 and the phage display library was constructed.Results The ScFv phage display library had a capacity of approximately 3.4×10~6,with 100% of the phagemids containing ScFv gene insertion as demonstrated by PCR.Conclusion The ScFv phage display library against human B-lymphoma was successfully constructed.The present study can be used for succeeding screening of specific antibody against human B-lymphoma.

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Available abstract

Objective To construct a single chain Fv(ScFv) phage display library against(human) B-lymphoma.Methods The mRNA was extracted from the spleen cells of the immunized(BALB/c) mice with Raji B-lymphoma cell strain and cDNA was synthesized by reverse transcriptase.The heavychain and light-chain variable region genes(V_H and V_L)of immunoglobulin were amplified by PCR and joined by a DNA linker to encode peptide as a single chain Fv(ScFv) fragment.ScFv fragments were amplified and cloned into the phagemid vector pHEN1 and the phage display library was constructed.Results The ScFv phage display library had a capacity of approximately 3.4×10~6,with 100% of the phagemids containing ScFv gene insertion as demonstrated by PCR.Conclusion The ScFv phage display library against human B-lymphoma was successfully constructed.The present study can be used for succeeding screening of specific antibody against human B-lymphoma.

Key concepts: Phage display, Phagemid, Raji cell, Molecular biology, Single-chain variable fragment, Antibody, Immunoglobulin light chain, Complementary DNA

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