3 years ago

Metastability Gap in the Phase Diagram of Monoclonal IgG Antibody

Metastability Gap in the Phase Diagram of Monoclonal IgG Antibody
Rachel A. Cancel, Ying Wang, Sergei Pechenov, Jacob B. Rowe, Rhiannon P. Flynn, Tyler D. Evangelous, Jifeng Zhang, J. Anand Subramony

Abstract

Crystallization of IgG antibodies has important applications in the fields of structural biology, biotechnology, and biopharmaceutics. However, a rational approach to crystallize antibodies is still lacking. In this work, we report a method to estimate the solubility of antibodies at various temperatures. We experimentally determined the full phase diagram of an IgG antibody. Using the full diagram, we examined the metastability gaps, i.e., the distance between the crystal solubility line and the liquid-liquid coexistence curve, of IgG antibodies. By comparing our results to the partial phase diagrams of other IgGs reported in literature, we found that IgG antibodies have similar metastability gaps. Thereby, we present an equation with two phenomenological parameters to predict the approximate location of the solubility line of IgG antibodies with respect to their liquid-liquid coexistence curves. We have previously shown that the coexistence curve of an antibody solution can be readily determined by the polyethylene glycol-induced liquid-liquid phase separation method. Combining the polyethylene glycol-induced liquid-liquid phase separation measurements and the phenomenological equation in this article, we provide a general and practical means to predict the thermodynamic conditions for crystallizing IgG antibodies in the solution environments of interest.

Publisher URL: http://www.cell.com/biophysj/fulltext/S0006-3495(17)30971-2

DOI: 10.1016/j.bpj.2017.08.048

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