Favourable interaction between heavy and light chains arrests the undesirable oligomerization of heavy chains in the refolding of denatured and reduced immunoglobulin G.
Recently we developed a slow dialysis method that effectively refolds denatured and reduced immunoglobulin G (IgG) [Maeda, Ueda and Imoto (1996) Prot. Engng 9: 95-100]. This method allows both individual and simultaneous refolding of denatured and reduced H and L chains. Analysis by SDS-polyacrylamide gel electrophoresis revealed that some oligomers were formed through disulfide bonds when H chains were refolded individually. It was also shown that the extent of IgG obtained by rejoining the mixture of refolded H and L chains which had been refolded individually was similar to that obtained by refolding denatured and reduced whole IgG. The results indicated that a favourable interaction between H and L chains prevented formation of H-chain oligomers to yield intact IgG. The present results suggest a mechanism whereby individually folded chains might associate to form IgG molecules in vivo.
Ueda
T
T
Graduate School of Pharmaceutical Sciences, Kyushu University 62, Fukuoka, Japan.
Maeda
Y
Y
So
T
T
Imoto
T
T
D000818:Animals; D000911:Antibodies, Monoclonal; D004220:Disulfides; D005978:Glutathione / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry*; D007147:Immunoglobulin Light Chains / Q000737:chemistry; D007142:Immunoglobulin gamma-Chains / Q000737:chemistry*; D046911:Macromolecular Substances; D008623:Mercaptoethanol / Q000737:chemistry; D051379:Mice; D010084:Oxidation-Reduction; D011489:Protein Denaturation; D017510:Protein Folding
D000911:Antibodies, Monoclonal; D004220:Disulfides; D007074:Immunoglobulin G; D007147:Immunoglobulin Light Chains; D007142:Immunoglobulin gamma-Chains; D046911:Macromolecular Substances; D008623:Mercaptoethanol; D005978:Glutathione