title: Expression, purification, and antibody binding activity of human papillomavirus 16 L1 protein fused to maltose binding protein.
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 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 HPV16 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 HPV16 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.
authors.affiliation: School of Life Sciences and Biotechnology, Korea University, Anam-dong, Seungbuk-gu, Seoul, South Korea.
authors.forename: Hee-Jeong
authors.initials: HJ
authors.lastname: Cho
authors.forename: Moon Sun
authors.initials: MS
authors.lastname: Hahm
authors.forename: Myung Kuk
authors.initials: MK
authors.lastname: Kim
authors.forename: In-Kwon
authors.initials: IK
authors.lastname: Han
authors.forename: Woon-Won
authors.initials: WW
authors.lastname: Jung
authors.forename: Han-Gon
authors.initials: HG
authors.lastname: Choi
authors.forename: Jung Ae
authors.initials: JA
authors.lastname: Kim
authors.forename: Yu-Kyoung
authors.initials: YK
authors.lastname: Oh
mesh_terms: D000914:Antibodies, Viral / Q000276:immunology; D000937:Antigen-Antibody Reactions; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry; D003001:Cloning, Molecular / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000737:chemistry; D052162:Human papillomavirus 16 / Q000737:chemistry*; D002479:Inclusion Bodies / Q000737:chemistry; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D058619:Maltose-Binding Proteins; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D012995:Solubility; D013696:Temperature
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D002352:Carrier Proteins; D058619:Maltose-Binding Proteins; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D007544:Isopropyl Thiogalactoside; C063317:L1 protein, Human papillomavirus type 16; D005982:Glutathione Transferase