Protein self-interaction is important in protein crystal growth, solubilization, and aggregation, both in vitro and in vivo, as with protein misfolding diseases, such as Alzheimer's. Although second virial coefficient studies can supply invaluable quantitative information, their emergence as a systematic approach to evaluating protein self-interaction has been slowed by the limitations of traditional measurement methods, such as static light scattering. Comparatively, self-interaction chromatography is an inexpensive, high-throughput method of evaluating the osmotic second virial coefficient (B) of proteins in solution. In this work, we used self-interaction chromatography to measure B of lysozyme in the presence of various cosolvents, including sucrose, trehalose, mannitol, glycine, arginine, and combinations of arginine and glutamic acid and arginine and sucrose in an effort to develop a better fundamental understanding of protein self-interaction in complex cosolvent systems. All of these cosolvents, alone or in combination, increased B, indicating a reduction in intermolecular attraction. However, the magnitude of cosolvent-induced changes in B was found to be largely dependent on the ability to control long-range electrostatic repulsion. To the best of our knowledge, this work represents the most comprehensive virial coefficient study to date focusing on complex cosolvent-induced effects on the self-interaction of lysozyme.