The Use of Combinatorial Heavy and Light Chain Libraries and Site Specific Mutagenesis to Create Antibody Biosensors for Metal Ions.
Abstract
A recursive protocol will be used to create the required Fab fragments. (1) Antibodies with the desired binding functions will be induced with the appropriate immunogen and Fabs obtained from recombinant libraries; (2) Fabs will be screened for antigen binding and those with appropriate affinities isolated, sequenced and overexpressed in E. coli; (3) The structures of selected Fabs will be deduced from modeling and amino acid motifs required for metal ion binding will be introduced by site specific mutagenesis; (4) purified Fabs will be tested for their metal ion affinity and response to a transducing substrate; (5) improvements in the binding and catalytic properties of the Fab will be sought by additional rounds of mutagenesis and/or chain shuffling with the original light or heavy chain libraries.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 31, 1992
- Accession Number
- ADA252020
Entities
People
- Stephen J. Benkovic
Organizations
- Pennsylvania State University