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- W2393834349 abstract "Objective: To resume the binding ability of a single chain Fv (ScFv) antibody against human VEGF by revising the point mutation and expressing it over again. Methods: From the hybridoma cell line secreting monoclonal antibody against VEGF, total RNA was prepared and used as a template for cDNA synthesis and cloning. Reverse transcription and polymerase chain reaction (RT-PCR) were used to clone the immunoglobulin variable region (V11H ,V11L ). The constructed ScFv was inserted into pET-15b with a E-tag at the 3'end. Compared with its commom and consensus sequence, the point mutations were confirmed and then revised by PCR. The binding activity of the revised ScFv was evaluated by ELJSA. Results: 4 point mutations were found in sequence of ScFv and the revised ScFv had a higher binding ability to VEGF 165 than mutated one. Conclusion: Based upon analyse of the conservative sequence of the amino residues and its property , it is a simple and convenient way to confirm the point mutations. The revised SvFv is potentially useful in both diagnostic and therapeutic application of human malignant tumors." @default.
- W2393834349 created "2016-06-24" @default.
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- W2393834349 date "2004-01-01" @default.
- W2393834349 modified "2023-09-26" @default.
- W2393834349 title "Cloning and reverse mutation of a single chain Fv antibody against human VEGF" @default.
- W2393834349 hasPublicationYear "2004" @default.
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