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Estimation of polyclonal IgG4 hybrids in normal human serum
Elizabeth Young1, Emma Lock, Douglas G Ward
1The Binding Site Group Ltd, Birmingham, UK.
Immunology
|February 12, 2014
Summary
Normal human serum contains significant levels of hybrid immunoglobulin G4 (IgG4) molecules, with both kappa and lambda light chains. These IgG4κ/λ hybrids constitute 21-33% of total IgG4 in healthy individuals.
Area of Science:
- Immunology
- Biochemistry
- Molecular Biology
Background:
- Hybrid immunoglobulin G4 (IgG4) molecules, formed by half-molecule exchange, have been observed experimentally.
- The prevalence of these hybrid IgG4 molecules in normal human serum is not well-established.
Purpose of the Study:
- To determine the incidence of polyclonal IgG4 hybrids in normal human serum.
- To investigate the presence of IgG4 molecules with differing immunoglobulin light chains.
Main Methods:
- Purification of polyclonal IgG4 from human sera.
- Fractionation using light-chain affinity and size exclusion chromatography.
- Analysis by SDS-PAGE, immunoblotting, ELISA, immunodiffusion, and mass spectrometry.
Main Results:
- Normal human serum IgG4 comprises IgG4κ, IgG4λ, and hybrid IgG4κ/λ molecules.
- Hybrid IgG4κ/λ molecules, composed of two IgG4 heavy chains and one kappa and one lambda light chain, ranged from 21% to 33% in analyzed sera.
- No hybrid IgG κ/λ antibodies were detected in IgG4-depleted polyclonal IgG.
Conclusions:
- Hybrid IgG4κ/λ antibodies represent a substantial fraction of total IgG4 in normal human serum.
- These findings highlight the complexity of IgG4 structure in physiological conditions.
