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Original article
Quantification of large and middle proteins of hepatitis B virus surface antigen (HBsAg) as a novel tool for the identification of inactive HBV carriers
  1. Maria Pfefferkorn1,
  2. Stephan Böhm2,
  3. Tina Schott1,
  4. Danilo Deichsel1,
  5. Corinna M Bremer3,
  6. Kathrin Schröder3,
  7. Wolfram H Gerlich3,
  8. Dieter Glebe3,
  9. Thomas Berg1,
  10. Florian van Bömmel1
  1. 1 Section of Hepatology, Clinic for Gastroenterology and Rheumatology, University Hospital Leipzig, Leipzig, Germany
  2. 2 Max von Pettenkofer—Institute for Hygiene and Clinical Microbiology, Ludwig-Maximilians-Universität, Munich, Germany
  3. 3 National Reference Center for Hepatitis B and D Viruses, Institute for Medical Virology, German Centre for Infection Research (DZIF), Justus Liebig University Giessen, Giessen, Germany
  1. Correspondence to Dr Florian van Bömmel, Section of Hepatology, Clinic for Gastroenterology and Rheumatology, University Hospital Leipzig AöR, Liebigstraße 20, 04103 Leipzig, Germany; florian.vanboemmel{at}


Objective Among individuals with chronic hepatitis B, those with hepatitis B e-antigen (HBeAg)-negative chronic hepatitis (CHB) can be difficult to distinguish from those with HBeAg-negative chronic HBV infection, also referred to as inactive HBV carriers (ICs), but both require different medical management. The level of HBV surface antigen (HBsAg) has been proposed as a marker to discriminate between chronic infection and hepatitis stages. HBsAg consists of large, middle and small HBs. The aim of this study was to determine whether the composition of HBsAg improved the identification of ICs among HBsAg-positive subjects with different phases of HBV infections.

Design HBV large surface proteins (LHBs) and HBV middle surface proteins (MHBs) were quantified in serum samples from 183 clinically well-characterised untreated patients with acute (n=14) HBV infection, ICs (n=44), CHBs (n=46), chronic HBeAg-positive phase (n=68) and hepatitis delta coinfection (n=11) using an ELISA, with well-defined monoclonal antibodies against the preS1 domain (LHBs) and the preS2-domain (MHBs). A Western blot analysis was used to verify the quantitation of the components of HBsAg. Total HBsAg was quantified using a modified commercially available assay (HBsAg V.6.0, Enzygnost, Siemens, Erlangen).

Results The composition of HBsAg showed specific patterns across different phases of hepatitis B. Individuals in the IC phase had significantly lower proportions of LHBs and MHBs than patients in acute or chronic phases irrespective of their HBV e-antigen status (p<0.0001) or HBsAg level. Both LHBs and MHBs ratios better predicted the IC phase than total HBsAg levels.

Conclusion Quantification of MHBs, particularly LHBs represents a novel tool for the identification of the IC stage.

  • HBsAg
  • LHBs
  • MHBs
  • preS1
  • preS2
  • subviral particles

Statistics from


  • Contributors MP designed the study, collected data, established and validated the ELISA, measured samples, analysed results and drafted the manuscript. SB contributed to establishing the ELISA. TS contributed to measuring HBsAg components. DD quantified HBV DNA and determined HBV genotypes. CMB and KS developed the WB for the HBsAg components and measured samples. TS, CMB, DD, KS, WHG, DG and TB critically revised the manuscript. FvB initiated, designed and supervised the study and drafted the manuscript. All authors have approved the final version for publication.

  • Funding Supported by Grant SFB 1021 B08 (to DG) from the Deutsche Forschungsgemeinschaft (DFG) and by grant no 934000-834 (DLR 01ES0821, to FvB and DG) from the German Ministry of Education and Research (BMBF). The National Reference Center for Hepatitis B and D Viruses at JLU Giessen is supported by the German Ministry of Health via the Robert Koch Institute, Berlin, Germany.

  • Competing interests None declared.

  • Patient consent Obtained.

  • Ethics approval Ethics Committees of Medical Research of the Universities of Leipzig and Berlin.

  • Provenance and peer review Not commissioned; externally peer reviewed.

  • Correction notice This article has been corrected since it published Online First. Figure 1 has been updated.

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