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Applied and Environmental Microbiology, November 2000, p. 5024-5029, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Specific Secretion of Active Single-Chain Fv Antibodies into the Supernatants of Escherichia coli Cultures by Use of the Hemolysin System

Luis A. Fernández, Isabel Sola, Luis Enjuanes, and Víctor de Lorenzo*

Centro Nacional de Biotecnología del Consejo Superior de Investigaciones Científicas (CSIC), Campus de Cantoblanco, 28049 Madrid, Spain

Received 12 May 2000/Accepted 22 August 2000

A simple method for the nontoxic, specific, and efficient secretion of active single-chain Fv antibodies (scFvs) into the supernatants of Escherichia coli cultures is reported. The method is based on the well-characterized hemolysin transport system (Hly) of E. coli that specifically secretes the target protein from the bacterial cytoplasm into the extracellular medium without a periplasmic intermediate. The culture media that accumulate these Hly-secreted scFv's can be used in a variety of immunoassays without purification. In addition, these culture supernatants are stable over long periods of time and can be handled basically as immune sera.

* Corresponding author. Mailing address: Centro Nacional de Biotecnología-CSIC, Campus de Cantoblanco, Madrid 28049, Spain. Phone: 34-91-585-45-36. Fax: 34-91-585-45-06. E-mail: vdlorenzo{at}cnb.uam.es.

Applied and Environmental Microbiology, November 2000, p. 5024-5029, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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