Expression, purification, and antibody binding activity of human papillomavirus 16 L1 protein fused to maltose binding protein
- Title
- Expression, purification, and antibody binding activity of human papillomavirus 16 L1 protein fused to maltose binding protein
- Author
- 최한곤
- Keywords
- ELISA; fusion protein; HPV16 L1 protein; maltose binding protein; solubilization
- Issue Date
- 2007-05
- Publisher
- BENTHAM SCIENCE PUBL LTD
- Citation
- PROTEIN AND PEPTIDE LETTERS, v. 14, No. 5, Page. 417-424
- 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.
- URI
- https://www.ingentaconnect.com/content/ben/ppl/2007/00000014/00000005/art00002https://repository.hanyang.ac.kr/handle/20.500.11754/106499
- ISSN
- 0929-8665; 1875-5305
- DOI
- 10.2174/092986607780782722
- Appears in Collections:
- COLLEGE OF PHARMACY[E](약학대학) > PHARMACY(약학과) > Articles
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