Effect of divalent ions on protein precipitation with polyethylene glycol: mechanism of action and applications
- PMID: 1821777
- DOI: 10.1016/1046-5928(91)90015-b
Effect of divalent ions on protein precipitation with polyethylene glycol: mechanism of action and applications
Abstract
Polyethylene glycol (PEG) is extensively employed for protein purification by fractional precipitation. Efficiency of precipitation is highest when the solution pH is near the isoelectric point of the target protein. At pH values far from the isoelectric point of the target protein, proteins develop a net positive or negative charge and are not more resistant to precipitation. We have found that divalent cations (Ba2+, Sr2+, and Ca2+) or divalent anions (SO4(2-)) significantly change the pattern of PEG precipitation when the ion is chosen so as to counteract the expected net charge on the target protein. At moderate (5-50 mM) concentrations of Ba2+, negatively charged proteins can be precipitated from solution at pH values as high as 10 with efficiency unchanged from precipitation at pH values near their isoelectric point values. The mechanism of PEG precipitation of protein at these high pH values appears to be unchanged from the mechanism operative at the protein isoelectric point. Precipitation is rapid and the capacity for protein precipitation is high. There is no detectable coprecipitation of small molecules (AMP, ATP, and NADH) or soluble proteins (carbonic anhydrase) induced when large quantities of protein are precipitated by this method. The purification of bovine carbonic anhydrase from erythrocyte lysate is more efficient at pH 10 in the presence of Ba2+ than is conventional PEG precipitation carried out at the isoelectric point of carbonic anhydrase. Application of these observations should broaden the utility of protein purification by fractional precipitation with PEG.
Similar articles
-
Limitations of polyethylene glycol-induced precipitation as predictive tool for protein solubility during formulation development.J Pharm Pharmacol. 2018 May;70(5):648-654. doi: 10.1111/jphp.12699. Epub 2017 Jan 20. J Pharm Pharmacol. 2018. PMID: 28106257
-
Large-scale purification of ovalbumin using polyethylene glycol precipitation and isoelectric precipitation.Poult Sci. 2019 Mar 1;98(3):1545-1550. doi: 10.3382/ps/pey402. Poult Sci. 2019. PMID: 30184130
-
A comparison of the dodecyl sulfate-induced precipitation of the myelin basic protein with other water-soluble proteins.Neurochem Res. 1986 Feb;11(2):299-315. doi: 10.1007/BF00967977. Neurochem Res. 1986. PMID: 3010147
-
Aqueous two-phase systems for protein separation: a perspective.J Chromatogr A. 2011 Dec 9;1218(49):8826-35. doi: 10.1016/j.chroma.2011.06.051. Epub 2011 Jun 21. J Chromatogr A. 2011. PMID: 21752387 Review.
-
Selective Precipitation of Proteins.Curr Protoc Protein Sci. 2016 Feb 2;83:4.5.1-4.5.37. doi: 10.1002/0471140864.ps0405s83. Curr Protoc Protein Sci. 2016. PMID: 26836410 Review.
Cited by
-
Continuous capture of recombinant antibodies by ZnCl 2 precipitation without polyethylene glycol.Eng Life Sci. 2020 Mar 24;20(7):265-274. doi: 10.1002/elsc.201900160. eCollection 2020 Jul. Eng Life Sci. 2020. PMID: 32647505 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous