Hepatitis C virus E2 protein purified from mammalian cells is frequently recognized by E2-specific antibodies in patient sera
SCIE
SCOPUS
- Title
- Hepatitis C virus E2 protein purified from mammalian cells is frequently recognized by E2-specific antibodies in patient sera
- Authors
- Lee, KJ; Suh, YA; Cho, YG; Cho, YS; Ha, GW; Chung, KH; Hwang, JH; Yun, YD; Lee, DS; Kim, CM; Sung, YC
- Date Issued
- 1997-11-28
- Publisher
- AMER SOC BIOCHEMISTRY MOLECULAR BIOLO
- Abstract
- 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 beta-mercapto-ethanol, 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).
- Keywords
- INSECT CELLS; NEUTRALIZING ANTIBODIES; ENVELOPE GLYCOPROTEINS; EXPRESSED PROTEIN; NON-A; GENOME; PURIFICATION; CLEAVAGE; E1; OLIGOMERIZATION
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/21162
- DOI
- 10.1074/jbc.272.48.30040
- ISSN
- 0021-9258
- Article Type
- Article
- Citation
- JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 272, no. 48, page. 30040 - 30046, 1997-11-28
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