Estimation of recombinant bovine somatotropin solubility by excluded-volume interaction with polyethylene glycols.
作者： Cynthia L. Stevenson , Michael J. Hageman
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摘要: Purpose. The potential to estimate protein solubilities, with limited protein, by excluded-volume interactions was evaluated using polyethylene glycols (PEG) and recombinant bovine Somatotropin (rbSt). Methods. Solutions of rbSt were prepared at concentrations significantly below saturation solubility. Subsequently, varying amounts PEG added force precipitation. Following centrifugation, the concentration in supernatant assayed reversed-phase HPLC, where a logarithmic relationship between solubility % observed. Results. An apparent absence determined extrapolation compared well values measured conventional approaches. Slopes log versus curves consistent principles depended on molecular weight used. Furthermore, precipitation process proved be reversible, allowing for recovery intact protein. Solubility-pH profiles obtained presence greatly reduced quantities needed favorably PEG. Conclusions. Thus, it appears feasible practical, certain limitations, obtain estimates proteins volume-exclusion methods supplies
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