Expression Studies of Catalytic Antibodies

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1995 Dec 5

PMID 8524873

Citations 14

Authors

H D Ulrich

P A Patten

P L Yang

F E Romesberg

P G Schultz

Affiliations

Soon will be listed here.

Abstract

We have examined the positive influence of human constant regions on the folding and bacterial expression of active soluble mouse immunoglobulin variable domains derived from a number of catalytic antibodies. Expression yields of eight hybridoma- and myeloma-derived chimeric Fab fragments are compared in both shake flasks and high density fermentations. In addition the usefulness of this system for the generation of in vivo expression libraries is examined by constructing and expressing combinations of heavy and light chain variable regions that were not selected as a pair during an immune response. A mutagenesis study of one of the recombinant catalytic Fab fragments reveals that single amino acid substitutions can have dramatic effects on the expression yield. This system should be generally applicable to the production of Fab fragments of catalytic and other hybridoma-derived antibodies for crystallographic and structure-function studies.

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