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Volume 272, Number 48, Issue of November 28, 1997 pp. 30040-30046
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Hepatitis C Virus E2 Protein Purified from Mammalian Cells Is Frequently Recognized by E2-specific Antibodies in Patient Sera

(Received for publication, July 1, 1997, and in revised form, September 18, 1997)

Ki Jeong Lee , Young-Ah Suh , Young Gyu Cho , Young Shik Cho ^¶ , Gun Woo Ha ^¶ , Kwang-Hoe Chung , Jae Hoon Hwang , Young Dae Yun , Dong Soon Lee ^** , Chang Min Kim ^** and Young-Chul Sung

From the  Department of Life Science, Center for Biofunctional Molecules, School of Environmental Engineering, Pohang University of Science and Technology, San 31, Hyoja Dong, Pohang, 790-784, ^¶ Korea Green Cross Corporation,  Mogam Biotechnology Research Institute, Yongin-City, Kyunggi-Do, and ^** Korea Cancer Center Hospital, Gongrung Dong, Nowonku, Seoul, Korea

The envelope protein of hepatitis C virus (HCV) is composed of two membrane-associated glycoproteins, E1 and E2. To obtain HCV E2 protein as a secretory form at a high level, we constructed a recombinant chinese hamster ovary (CHO) cell line expressing a C-terminal truncated E2 (E2t) fused to human growth hormone (hGH), CHO/hGHE2t. The hGHE2t fusion protein was purified from the culture supernatant using anti-hGH mAb affinity chromatography at approximately 80% purity. The purified hGHE2t protein appeared to be assembled into oligomers linked by intermolecular disulfide bond(s) when density gradient centrifugation and SDS-polyacrylamide gel electrophoresis were employed. When the purified fusion protein was used for testing its ability to bind to antibodies specific for HCV by enzyme-linked immunosorbent assay, the protein was recognized by antibodies in sera from 90% of HCV-positive patients. Treatment of hGHE2t protein by -mercaptoethanol, but not by heat and SDS, significantly reduced its reactivity to the antibodies of patient sera, suggesting that intermolecular and/or intramolecular disulfide bonds are important for its ability to recognize its specific antibody and that the E2 protein contains discontinuous antigenic epitope(s).

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