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篇目详细内容 |
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【篇名】 |
Preparation, identification, and clinical application of anti-HBs monoclonal antibody that binds both wild-type and immune escape mutant HBsAgs |
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【刊名】 |
Frontiers of Medicine in China |
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【刊名缩写】 |
Front. Med. China |
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【ISSN】 |
1673-7342 |
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【EISSN】 |
1673-7458 |
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【DOI】 |
10.1007/s11684-009-0047-0 |
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【出版社】 |
Higher Education Press and Springer-Verlag |
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【出版年】 |
2009 |
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【卷期】 |
3
卷3期 |
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【页码】 |
277-283
页,共
7
页 |
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【作者】 |
Fanghe LI BM;
Chunyan ZHANG MS;
Jinghua LIU MS;
Xiaoyan ZHANG MS;
Bing YAN;
Bo ZHANG MS;
Yongguo HUANG MS;
Jingsong GONG BM;
Yan CHEN BM;
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【关键词】 |
hepatitis B virus; gene variation; hepatitis B surface antigen; immune escape; enzyme-linked immunosorbent assay |
【摘要】 |
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Using a standard cellular fusion technique and indirect enzyme-linked immunosorbent assay (ELISA), a hybridoma cell line strain secreting anti-HBs monoclonal antibody (mAb) (defined G6 mAb) was obtained. The cells grew and secreted mAb stably. Antibody titers in the culture supernatant and ascites were 2.048×106 and 4.096×106, respectively. By applying the anti-HBs G6 mAb and horseradish peroxidase (HRP)-labeled goat anti-HBs antibody, we developed a sandwich ELISA (defined G6m ELISA) for detecting both wild-type and immune escape mutant HBsAgs (IEM HBsAg). The assay was performed to detect 17 species of genome recombinant expression HBsAg, including two wild-type species and 15 IEM HBsAg species, which varied in the “a” determinant, in a group of patients infected with hepatitis B virus (HBV). The patients previously had a lower ELISA detection signal [(absorbance of patients/absorbance of normal people (P/N): 1.0─4.5)]. The results demonstrated that the sensitivity of this assay to wild-type HBsAg was no less than 0.125g/L; 12 of 15 IEM HBsAg species (P/N≥2.5) were positive for G6 mAb. Of the positive IEM HBsAg species, two had a low absorbance value at 450nm (A450), one had an intermediate A450 value and nine had a high A450 value, which was 7.55%(mean), 59.4% and 92.1%─109.4% of the wild-type A450 value, respectively. The two species with low A450 value and the three negative species mutated at the bases 120─124 in the first loop of the HBV “a” determinant. Using the G6 ELISA and two commercial ELISA kits (A and B), 177 patients were tested. The G6 ELISA had a significantly higher detection rate than either commercial ELISAs (19.21% vs 14.89% and 6.21%, respectively; P<0.01, P<0.05, respectively). |
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