Precipitability and composition of HBsAg-anti-HBs immune complexes formed in the presence of complement. A model of circulating immune complex analysis

N. Anh-Tuan, A. Falus, G. Füst, Katalin Merétey, G. A. Medgyesi, S. Hollán

Research output: Contribution to journalArticle

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Abstract

Immune complexes (IC) of partially purified HBsAg and human anti-HBs were prepared at different antigen/antibody ratios in the presence of complement in normal human serum (NHS), and under conditions not allowing complement activation in buffers or in NHS containing 10 mM EDTA (NHS-EDTA). Commercial preparations of the radiolabelled antigen and antibody were used. IC formed in NHS were not significantly precipitated even after incubation for 24 h at 4°C, whereas a typical precipitation curve was observed with complexes formed in the absence of complement. Thus, complement activation was found to markedly and permanently inhibit precipitability of HBsAg-anti-HBs immune complexes (HBsAg-IC). HBsAg-IC were precipitated from sera with 3.5% polyethylene glycol (PEG), boiled in sodium dodecyl sulphate (SDS)-urea buffer, and analysed by SDS-polyacrylamide slab gel electrophoresis (SDS-PAGE). With complexes formed in the presence of complement, about one-sixth of the antibody activity was found in high molecular weight fractions corresponding in size to IgG oligomers. By contrast, with complexes formed without complement, no significant amount of antibody was found in these fractions. With blotting technique and radiolabelled anti-human-C3 antibody, it was demonstrated that anti-HBs was covalently bound to C3b fragments in IC formed in the presence of complement and was in the high molecular weight fractions.

Original languageEnglish
Pages (from-to)283-294
Number of pages12
JournalJournal of Immunological Methods
Volume64
Issue number3
DOIs
Publication statusPublished - Nov 25 1983

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Hepatitis B Surface Antigens
Antigen-Antibody Complex
Antibodies
Serum
Complement Activation
Edetic Acid
Sodium Dodecyl Sulfate
Buffers
Molecular Weight
Antigens
Urea
Polyacrylamide Gel Electrophoresis
Immunoglobulin G

Keywords

  • complement solubilization of complexes
  • precipitability of immune complexes
  • soluble immune complexes

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Precipitability and composition of HBsAg-anti-HBs immune complexes formed in the presence of complement. A model of circulating immune complex analysis. / Anh-Tuan, N.; Falus, A.; Füst, G.; Merétey, Katalin; Medgyesi, G. A.; Hollán, S.

In: Journal of Immunological Methods, Vol. 64, No. 3, 25.11.1983, p. 283-294.

Research output: Contribution to journalArticle

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AU - Hollán, S.

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