title: Size and structure of antigen-antibody complexes: thermodynamic parameters.
abstract: The role of antigen-antibody (Ag-Ab) complexes in the immune response depends, in part, on the size of the complexes. Previously, we combined electron microscopy with classical and quasi-elastic light scattering to characterize the molecular weight distribution and the conformation of Ag-Ab complexes made from bovine serum albumin (BSA) and pairs of anti-BSA monoclonal antibodies at a single concentration and Ag:Ab molar ratio. In this report, the molecular weight distribution of Ag-Ab complexes was determined by classical light scattering at a single Ag:Ab ratio and over a range of concentrations, and binding of BSA to pairs of MAb was determined by radioimmunoassay at several Ag:Ab molar ratios. A thermodynamic model was developed for the equilibrium size distribution of Ag-Ab complexes formed between a pair of MAb, each with unique affinity and specificity, and an Ag containing a single epitope for each of the pair of MAb. The combined experimental data were used in conjunction with the model to determine the values of cyclization and polymerization constants. Successful determination of the parameters required data from both classical light scattering and electron microscopy. Cyclization constants were lower than those reported in other studies of Ag-Ab complexes; this may be attributable to our use of a protein Ag, as compared to a divalent hapten. In two out of three cases, cyclization constants increased with increasing number of Ab in the complex, in contrast to previous assumptions. The validity of the thermodynamic model was further shown by its ability, in combination with conformational and hydrodynamic model, to predict the hydrodynamic radius of the complexes over a wide range of experimental conditions.
authors: Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge 02139.
authors: R M
authors: RM
authors: Murphy
authors: R A
authors: RA
authors: Chamberlin
authors: P
authors: P
authors: Schurtenberger
authors: C K
authors: CK
authors: Colton
authors: M L
authors: ML
authors: Yarmush
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000915:Antibody Affinity; D000936:Antigen-Antibody Complex / Q000737:chemistry*; D007136:Immunoglobulins / Q000737:chemistry / Q000648:ultrastructure; D066298:In Vitro Techniques; D008027:Light; D046911:Macromolecular Substances; D051379:Mice; D011487:Protein Conformation; D012542:Scattering, Radiation; D012710:Serum Albumin, Bovine / Q000276:immunology; D013816:Thermodynamics
chemical_list: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007136:Immunoglobulins; D046911:Macromolecular Substances; D012710:Serum Albumin, Bovine