Expression, purification, and antibody binding activity of human papillomavirus 16 L1 protein fused to maltose binding protein
- Authors
- Cho, Hee-Jeong; Hahm, Moon Sun; Kim, Myung Kuk; Han, In-Kwon; Jung, Woon-Won; Choi, Han-Gon; Kim, Jung Ae; Oh, Yu-Kyoung
- Issue Date
- Mar-2007
- Publisher
- BENTHAM SCIENCE PUBL LTD
- Keywords
- HPV16 L1 protein; fusion protein; maltose binding protein; solubilization; ELISA
- Citation
- PROTEIN AND PEPTIDE LETTERS, v.14, no.5, pp.417 - 424
- Indexed
- SCIE
SCOPUS
- Journal Title
- PROTEIN AND PEPTIDE LETTERS
- Volume
- 14
- Number
- 5
- Start Page
- 417
- End Page
- 424
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/44377
- DOI
- 10.2174/092986607780782722
- ISSN
- 0929-8665
- Abstract
- Genetic human papillornavirus 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, HPV 16 L1 protein, which is required as a coating protein, has previously been expressed from costly and laborious recombinant baculovirus-infected 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 HPV 16 L1 protein (MBP-HPV16 L1) was 35% at 37 degrees C, but increased to 85% at 22 degrees 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 HPV 16 L1 under optimized temperature conditions, and that MBP-fused HPV16 L1 might be applied further in evaluations of the immune responses of HPV16 L1-based cervical cancer vaccines.
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