Accession Number:

ADA252020

Title:

The Use of Combinatorial Heavy and Light Chain Libraries and Site Specific Mutagenesis to Create Antibody Biosensors for Metal Ions.

Descriptive Note:

Annual rept. 1 Jun 91-31 May 92,

Corporate Author:

PENNSYLVANIA STATE UNIV UNIVERSITY PARK

Personal Author(s):

Report Date:

1992-05-31

Pagination or Media Count:

3.0

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 andor chain shuffling with the original light or heavy chain libraries.

Subject Categories:

  • Medicine and Medical Research
  • Microbiology

Distribution Statement:

APPROVED FOR PUBLIC RELEASE