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Functional expression of a single-chain antibody specific for the HER2 human oncogene in a bacterial reducing environment

TitleFunctional expression of a single-chain antibody specific for the HER2 human oncogene in a bacterial reducing environment
Publication TypeArticolo su Rivista peer-reviewed
Year of Publication2005
AuthorsLombardi, Alessio, Sperandei Maria, Cantale Cristina, Giacomini Patrizio, and Galeffi Patrizia
JournalProtein Expression and Purification
Volume44
Pagination10 - 15
Date Published2005///
KeywordsCytoplasmic antibodies, HER2, Single-chain fragment of variable antibody regions
Abstract

Recombinant antibody fragments represent useful tools for cancer diagnosis and therapy. Aberrant expression of the HER2 receptor is implicated in metastatic breast and ovary cancers, two malignancies with a high prevalence in young women. In this study, we focussed on a single-chain fragment of variable antibody regions specific for HER2 (scFv800E6) that can be expressed in a functional form in the cytoplasm of Escherichia coli. ScFv800E6 was extracted from bacterial cultures following induction at different temperatures and purified. The yield of both soluble and insoluble forms was measured. We found that scFv800E6 was functional when expressed in the soluble fraction in the bacteria cytosol. In addition, scFv800E6 extracted from inclusion bodies was easily refolded and largely recovered its functionality. Thus, scFv800E6 is intrinsically capable of efficient and functional folding in a reducing environment and represents one of the few described antibody fragments with a framework well adapted for cytoplasmic expression. © 2005 Elsevier Inc. All rights reserved.

Notes

Cited By (since 1996): 6Export Date: 25 August 2010Source: Scopus

URLhttp://www.scopus.com/inward/record.url?eid=2-s2.0-26044461188&partnerID=40&md5=6d982ead4969bc64acb9685745e73a16
Citation Key275