Selection and characterization of lipase abzyme from phage displayed antibody libraries |
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Authors: | Leong Max K Chen Chinpiao Shar Ker-Chang Shiuan David |
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Institution: | Department of Chemistry, National Dong Hwa University, Hualien 974, Taiwan, ROC. |
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Abstract: | Antibodies with enzymatic activity were named abzymes or catalytic antibodies. In the present study, the lipolytic abzymes were selected from the phage displayed antibody libraries against a transition state analog (TSA) of lipases/esterases. After three rounds of selection, four monoclonal phage particles capable of binding significantly with the TSA were obtained. The soluble scFv antibody fragments were further expressed and obtained using Escherichia coli strain HB2151. The binding capabilities and the apparent enzymatic activities of the purified antibody proteins were measured. The 3D structures of the expressed antibodies were also predicted through homology modeling and binding-site prediction algorithm. The present method demonstrates that selection from phage displayed antibody libraries is an efficient and convenient means to find new abzymes. |
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Keywords: | PBS phosphate buffered saline solution scFv single-chain variable regions of antibodies TSA transition state analog |
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