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Expression of human cytotoxic T-lymphocyte antigen 4 (hCTLA4) fused with Hepatitis B surface antigen (HBsAg) using Pichia pastoris

Paper ID Volume ID Publish Year Pages File Format Full-Text
18446 42723 2006 5 PDF Available
Title
Expression of human cytotoxic T-lymphocyte antigen 4 (hCTLA4) fused with Hepatitis B surface antigen (HBsAg) using Pichia pastoris
Abstract

Human cytotoxic T-lymphocyte antigen 4 (hCTLA4), an immunosuppressive agent, was expressed as a fusion protein to produce a multimeric form. The fusion partner was Hepatitis B surface antigen (HBsAg), which was fused to the N-terminal extracellular domain of hCTLA4. The expression vector, pPIC3.5hc, constructed for the insertion of the fusion gene into the pPIC3.5 vector with the AOX promoter, was introduced into Pichia pastoris GS115. The expressed fusion protein, HBsAg–hCTLA4, had a molecular size of ∼39 kDa, which was detected using anti-hCTLA4 and anti-HBsAg antibodies by Western blot analysis. Transmission electron microscopic analysis of the expressed fusion protein revealed two kinds of particles including 40 nm fused particles and 22 nm HBsAg particles. The 40 nm particles were assumed to be particles fused with hCTLA4 displayed as a multimeric form on the outer surface of the HBsAg particles. The HBsAg in the HBsAg–hCTLA4 particles were shown to have a low antigenicity in reversed passive hemagglutination assay (RPHA). Accordingly, the HBsAg was proven to be an effective fusion partner. In addition, hCTLA4 fused with HBsAg could possibly be displayed as a multimeric form.

Keywords
Human cytotoxic T-lymphocyte antigen 4; Hepatitis B surface antigen; Pichia pastoris
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Expression of human cytotoxic T-lymphocyte antigen 4 (hCTLA4) fused with Hepatitis B surface antigen (HBsAg) using Pichia pastoris
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 39, Issue 3, 3 July 2006, Pages 481–485
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us