DIRECT MEASUREMENT VIA PHAGE TITER OF THE DISSOCIATION-CONSTANTS IN SOLUTION OF

2020-03-26 15:16:30

protein binding phage Edinburgh

责任者: DYSON, MR;GERMASCHEWSKI, V;MURRAY, K 单位: UNIV EDINBURGH,INST CELL & MOLEC BIOL,EDINBURGH EH9 3JR,MIDLOTHIAN,SCOTLAND. 来源出处: NUCLEIC ACIDS RESEARCH, v 23, MAY 11 1995, p 1531- 1535 摘要: Studies of interactions between filamentous fusion phage particles and protein or nucleic acid molecules have gained increasing importance with recent successes of screening techniques based upon random phage display libraries (biopanning). Since a number of different phage are usually obtained by biopanning, it is useful to compare quantitatively the binding affinities of individual phage for the substrate used for selection. A procedure is described for determination of relative dissociation constants (K-d(Rel)) between filamentous phage carrying peptide fusions to the coat protein gpIII and substrates in solution. This novel method is based on the measurement of phage titres. Phage selected from a random fusion phage library for binding to a monoclonal antibody or a viral structural protein exhibited K-d(Rel) values in the nanomolar and micromolar ranges for their respective substrates, thus validating the method over a wide range of binding affinities. 关键词: LEAD; PROTEIN KINASE C; EXOCYTOSIS; CHROMAFFIN CELLS; NOREPINEPHRINE; HEPATITIS-B VIRUS; PEPTIDE LIBRARIES; FILAMENTOUS PHAGE; EPITOPE LIBRARY; BINDING; SELECTION; PROTEINS; ANTIBODY; ANTIGEN; IDENTIFICATION LIBRARY; BINDING; SELECTION; PROTEINS; ANTIBODY; ANTIGEN; IDENTIFICATION