Estimation of recombinant bovine somatotropin solubility by excluded-volume interaction with polyethylene glycols.
OBJECTIVE
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 of PEG were added to force protein precipitation. Following centrifugation, the protein concentration in the supernatant was assayed by reversed-phase HPLC, where a logarithmic relationship between solubility and % PEG was observed.


RESULTS
An apparent protein solubility in the absence of PEG was determined by extrapolation and compared well with values measured by conventional approaches. Slopes of log solubility versus % PEG curves were consistent with excluded-volume principles and depended on the molecular weight of the PEG used. Furthermore, the precipitation process proved to be reversible, allowing for recovery of intact protein. Solubility-pH profiles obtained in the presence of PEG greatly reduced the quantities of protein needed and compared favorably with profiles in the absence of PEG.


CONCLUSIONS
Thus, it appears feasible and practical, with certain limitations, to obtain solubility estimates of proteins by volume-exclusion methods with limited supplies of protein.
Stevenson
C L
CL
Alza Corporation, Palo Alto, California 94303, USA.
Hageman
M J
MJ
D000818:Animals; D002021:Buffers; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D013006:Growth Hormone / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D012995:Solubility
D002021:Buffers; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D013006:Growth Hormone