Accession Number:

ADA277000

Title:

Genetic Selection for Improved Abzymes in E. Coli

Descriptive Note:

Final rept. 1 Sep 1990-31 Jul 1993

Corporate Author:

SCRIPPS RESEARCH INST LA JOLLA CA

Personal Author(s):

Report Date:

1994-08-03

Pagination or Media Count:

5.0

Abstract:

In analogy to the evolutionary processes that have brought natural enzymes to peak efficiency, we are developing molecular selection processes in the laboratory for preparing and improving antibodies possessing catalytic activity abzymes. Over the three-year course of this project, we have achieved several important goals. We have exploited phage display methods for the first time to engineer functional single chain versions of an antibody with chorismate mutase activity, selecting for optimal linkers to join the relevant VH and VL domains. We have also established a sensitive growth selection assay in E. coli for catalysts with chorismate mutase activity, allowing for the direct selection for improved variants of our first-generation abzymes. Finally, we have utilized a sensitive immunoassay to screen antibody phage libraries directly for catalysis of a bimolecular Diels-Alder reaction and have identified several active clones. Detailed characterization of these molecules will allow the efficacy of standard hybridoma techniques and phage display methods to be directly compared. Catalytic antibody, Transition state analog, Cloning, Phage display, Genetic selection.

Subject Categories:

  • Biology
  • Medicine and Medical Research
  • Microbiology

Distribution Statement:

APPROVED FOR PUBLIC RELEASE