Preparation and biologic characterization of fragments containing dimeric and monomeric C gamma 2 domain of rabbit IgG.
A number of fragments derived from acid-treated rabbit IgG by digestion with plasmin have been separated by high-resolution gel filtration. Fragments isolated included a dimer and monomer Facb, named F(acb)2 and Facb, respectively and a heterodimer composed of Facb and Fab subunits, named F(acb)(ab). A C gamma 2 fragment was obtained by papain digestion of Facb. A heterodimer composed of Facb and Fab', named F(acb)(ab'), was also prepared by oxidizing a reduced mixture of these fragments. Fragments thus obtained are classified into two groups--those carrying paired C gamma 2 domains, i.e. F(acb)2, and the disulfide-linked dimeric C gamma 2 fragment; and those having a single C gamma 2 domain, i.e. reduced, alkylated Facb and C gamma 2 fragment, F(acb)(ab) and F(acb)(ab'). These fragments exhibited marked differences in their capacity to activate complement in assay systems of hemolysis and complement consumption by immune complexes or aggregates on polystyrene latex. Fragments of the former group could activate complement but with a definitely reduced efficiency (50%) compared to intact IgG, whereas fragments of the latter group were practically inactive. Although it was not determined whether the C1-binding capacity itself is changed by monomerization of the C gamma 2 domain, the results suggested that the intact paired C gamma 2 module is required at least for the activation process of complement.
D000818:Animals; D000936:Antigen-Antibody Complex / Q000276:immunology; D001704:Biopolymers; D002850:Chromatography, Gel; D003167:Complement Activation; D004591:Electrophoresis, Polyacrylamide Gel; D006461:Hemolysis; D007140:Immunoglobulin Fab Fragments / Q000032:analysis; D007141:Immunoglobulin Fc Fragments / Q000032:analysis / Q000302:isolation & purification; D007128:Immunoglobulin Fragments / Q000032:analysis* / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000032:analysis*; D007700:Kinetics; D008970:Molecular Weight; D011817:Rabbits
D000936:Antigen-Antibody Complex; D001704:Biopolymers; D007140:Immunoglobulin Fab Fragments; D007141:Immunoglobulin Fc Fragments; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; C041323:immunoglobulin Facb fragment