Expression, Purification, and Antibody Binding Activity of Human Papillomavirus 16 L1 Protein Fused to Maltose Binding Protein

Author(s): Hee-Jeong Cho , Moon Sun Hahm , Myung Kuk Kim , In-Kwon Han , Woon-Won Jung , Han-Gon Choi , Jung Ae Kim , Yu-Kyoung Oh .

Journal Name: Protein & Peptide Letters

Volume 14 , Issue 5 , 2007

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Abstract:

Genetic human papillomavirus type 16 L1 (HPV16 L1) has been widely studied for cervical cancer vaccine development. For the enzyme-linked immunosorbent assay (ELISA) screening of these vaccines, HPV16 L1 protein, which is required as a coating protein, has previously been expressed from costly and laborious recombinant baculovirusinfected insect cells. For a novel HPV16 L1 expression system characterized by a high yield of soluble form with simple purification steps, we have cloned and expressed two different types of HPV16 L1, both fused to maltose binding protein (MBP) or glutathione-S-transferase (GST) in Escherichia coli. The yield of soluble HPV16 L1 was influenced by the cultivation temperature. The yield of soluble form in the total MBP-fused HPV16 L1 protein (MBP-HPV16 L1) was 35% at 37°C, but increased to 85% at 22°C. Among the fusion partners, MBP provided higher yields of total and soluble HPV16 L1 than did GST. MBP-HPV16 L1 showed a 4.9-fold higher yield of the soluble form over insoluble inclusion bodies under optimized culture conditions. The soluble form of MBP-HPV16 L1 was purified via MBP affinity chromatography in a recovery yield of 9.7%. After fusion with MBP, HPV16 L1 showed binding activity to HPV16 L1-specific monoclonal antibody comparable to HPV16 L1 from the insect cells in ELISA tests. These results demonstrate that the use of MBP as a fusion partner may generate a high yield of soluble HPV16 L1 under optimized temperature conditions, and that MBPfused HPV16 L1 might be applied further in evaluations of the immune responses of HPV16 L1-based cervical cancer vaccines.

Keywords: hpv protein, fusion protein, maltose binding protein, solubilization, elisa

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Article Details

VOLUME: 14
ISSUE: 5
Year: 2007
Page: [417 - 424]
Pages: 8
DOI: 10.2174/092986607780782722
Price: $58

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