Forms of a self associating autoantibody complex between a monoclonal human IgG1 and human serum albumin

The mode of association of an unusual human autoantibody complex, composed of a monoclonal immunoglobulin, Tu IgG, and human serum albumin was investigated. A crystalline complex forms from these components in the cold and we have shown that it consists of IgG and albumin in a 1:2 molar ratio [Jentoft et al., Biochemistry 21, 289-294 (1982)]. The crystalline complex was analyzed by electron microscopy and the soluble natural complexes (formed by dissolving the crystals at 20 degrees C) were studied by sedimentation velocity. The sedimentation studies demonstrated that the soluble Tu IgG-albumin complexes are in equilibrium with free Tu IgG and albumin molecules and that the major soluble sedimenting species has a S20,w value of 12.5S. At a constant concn of complex, the size of the sedimenting complex can be reduced by lowering the pH, increasing the ionic strength, or adding CaCl2, citrate, ascorbate or urea. These intermediate, soluble forms have S20,w values that are consistent with 1:1 and 1:2 Tu IgG-albumin complexes. Parameters of repeat distances and angles that were obtained from electron micrographs of the crystalline form of the Tu IgG-albumin complex were used to propose a model for the 12.5S species and were also incorporated into a three-dimensional model for the complex. The 12.5S complex is proposed to form by dimerization of the 1:2 Tu IgG-albumin complex via interactions of albumin with the Fc region of the antibody. The 12.5S dimer may be the nucleating species for subsequent rapid associations that lead to spontaneous formation of crystals. In the proposed model for the Tu IgG-albumin crystals, the angle between the Fab arms of each Tu IgG molecule is 90 degrees, the antigenic determinant on the albumin is located near one end of the long axis of the cylindrical molecule, the site of interaction with Fc is located at the other end of the cylinder, and the CH3 domain of the IgG contains the binding site for albumin that is responsible for the formation of the dimeric 12.5S species. A series of sedimentation velocity experiments suggest that the association between the CH3 domain of IgG and albumin requires the prior formation of the antibody-antigen complex.