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Efficient production of complement (C3d)3 fusion proteins using the baculovirus expression vector system

Lookup NU author(s): Dr Andrew Knight

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Abstract

Proteins fused to activated complement (C) fragments elicit enhanced immunogenicity. This "natural adjuvant" effect may have important implications when considering novel vaccination approaches. Here we describe both the construction of a novel fusion protein, consisting of a well characterized test antigen fused to multiple copies of the activated complement component (C3d)3, as well as an efficient method for its expression and production in insect cells. Using the inherent biological advantages of the baculovirus expression system, as well as applying specific infection and harvesting modifications, we have optimized the efficiency of protein production. Our modifications allow purification of fusion proteins directly from cell supernatant in a single anion exchange chromatographic step. This alleviates the requirement for the inclusion of protein affinity tags. The integrity of the purified recombinant protein was evaluated by SDS PAGE analysis, reactivity with antibodies, as well as in vivo by administration as an immunogen. © 2005 Elsevier B.V. All rights reserved.

Publication metadata

Author(s): Barrault DV, Steward M, Cox VF, Smith RAG, Knight AM

Publication type: Article

Publication status: Published

Journal: Journal of Immunological Methods

Year: 2005

Volume: 304

Issue: 1-2

Pages: 158-173

Print publication date: 01/09/2005

ISSN (print): 0022-1759

ISSN (electronic): 1872-7905

Publisher: Elsevier

URL: http://dx.doi.org/10.1016/j.jim.2005.07.013

DOI: 10.1016/j.jim.2005.07.013

PubMed id: 16109422

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