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Article: Preparation, identification, and clinical application of anti-HBs monoclonal antibody that binds both wild-type and immune escape mutant HBsAgs

TitlePreparation, identification, and clinical application of anti-HBs monoclonal antibody that binds both wild-type and immune escape mutant HBsAgs
Authors
KeywordsEnzyme-Linked Immunosorbent Assay
Gene Variation
Hepatitis B Surface Antigen
Hepatitis B Virus
Immune Escape
Issue Date2009
PublisherGaodeng Jiaoyu Chubanshe. The Journal's web site is located at http://www.springer.com/medicine/journal/11684
Citation
Frontiers of Medicine in China, 2009, v. 3 n. 3, p. 277-283 How to Cite?
AbstractUsing 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 0.125 μg/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 450 nm (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). © Higher Education Press and Springer-Verlag GmbH 2009.
Persistent Identifierhttp://hdl.handle.net/10722/92213
ISSN
References

 

DC FieldValueLanguage
dc.contributor.authorLi, Fen_HK
dc.contributor.authorZhang, Cen_HK
dc.contributor.authorLiu, Jen_HK
dc.contributor.authorZhang, Xen_HK
dc.contributor.authorYan, Ben_HK
dc.contributor.authorZhang, Ben_HK
dc.contributor.authorHuang, Yen_HK
dc.contributor.authorGong, Jen_HK
dc.contributor.authorChen, Yen_HK
dc.date.accessioned2010-09-17T10:39:24Z-
dc.date.available2010-09-17T10:39:24Z-
dc.date.issued2009en_HK
dc.identifier.citationFrontiers of Medicine in China, 2009, v. 3 n. 3, p. 277-283en_HK
dc.identifier.issn1673-7342en_HK
dc.identifier.urihttp://hdl.handle.net/10722/92213-
dc.description.abstractUsing 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 0.125 μg/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 450 nm (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). © Higher Education Press and Springer-Verlag GmbH 2009.en_HK
dc.languageengen_HK
dc.publisherGaodeng Jiaoyu Chubanshe. The Journal's web site is located at http://www.springer.com/medicine/journal/11684en_HK
dc.relation.ispartofFrontiers of Medicine in Chinaen_HK
dc.subjectEnzyme-Linked Immunosorbent Assayen_HK
dc.subjectGene Variationen_HK
dc.subjectHepatitis B Surface Antigenen_HK
dc.subjectHepatitis B Virusen_HK
dc.subjectImmune Escapeen_HK
dc.titlePreparation, identification, and clinical application of anti-HBs monoclonal antibody that binds both wild-type and immune escape mutant HBsAgsen_HK
dc.typeArticleen_HK
dc.identifier.emailChen, Y:ychenc@hkucc.hku.hken_HK
dc.identifier.authorityChen, Y=rp1318en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1007/s11684-009-0047-0en_HK
dc.identifier.scopuseid_2-s2.0-69549114426en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-69549114426&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume3en_HK
dc.identifier.issue3en_HK
dc.identifier.spage277en_HK
dc.identifier.epage283en_HK
dc.identifier.eissn1673-7458-
dc.identifier.citeulike5127316-

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