Versatile E. coli thioredoxin specific monoclonal antibodies afford convenient analysis and purification of prokaryote expressed soluble fusion protein

Richard R. Dickason, Robert A. Edwards, Joseph Bryan, David P. Huston

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

A recently developed E. coli thioredoxin (Trx) gene fusion expression system has circumvented the difficulties associated with inclusion body formation. Although ample quantities of soluble recombinant protein can be expressed using this system, no universal means of quantifying or purifying the fusion product exists. To facilitate the study of Trx fusion proteins, anti-E. coli Trx monoclonal antibodies (mAb) were generated. Two distinct Trx epitopes were defined by competitive ELISA. Both mAb were capable of detecting Trx fusion proteins by sandwich ELISA, and by immunoblot analysis under reducing and non-reducing conditions. In addition, these mAb enabled purification of Trx fusion proteins by immunoprecipitation, as well as affinity chromatography. This report provides the first description of anti-Trx antibodies. These reagents represent a major advance in the isolation and analysis of prokaryote expressed recombinant Trx fusion proteins.

Original languageEnglish (US)
Pages (from-to)237-244
Number of pages8
JournalJournal of Immunological Methods
Volume185
Issue number2
DOIs
StatePublished - 1995

Keywords

  • Chromatography affinity
  • ELISA
  • Immunoblot analysis
  • Monoclonal antibody
  • Recombinant protein
  • Thioredoxin

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Biotechnology

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