Conditions for the adsorption of proteins on ultrastable zeolite Y and its use in protein purification.
The adsorption of proteins on ultrastable zeolites was investigated. Protein binding to one of these, ultrastable zeolite Y (USY), was studied in detail. Protein binding to USY, with a Si/Al ratio of > 240, was found to be dependent on the pH of the solution, being highest at or just below the pI of the protein. The amount of protein adsorbed on the zeolite was found to be 10 times as much as the estimated binding to the external surface of the USY. We propose an adsorption mechanism involving the formation of a protein layer strongly bound to the USY surface, further protein layers being formed on top of this on the basis of protein-protein interactions. The protein-protein interactions can be disrupted by changing the pH. Ultrastable zeolite Y was used as a new matrix for protein purification. Undesired proteins can be removed from a crude preparation by adsorption on USY, increasing the purity of a specific protein, or the protein can be adsorbed on the zeolite and subsequently eluted through changing the pH. These two means of protein purification are exemplified by the purification of peroxidase from a crude horseradish extract and by the purification of lysozyme from egg white.
Klint
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Department of Inorganic Chemistry 2, Lund University, Sweden.
Eriksson
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H
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002645:Chickens; D006735:Horseradish Peroxidase / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D009113:Muramidase / Q000302:isolation & purification / Q000378:metabolism; D010047:Ovalbumin / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000493:pharmacokinetics*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification / Q000378:metabolism; D014462:Ultrafiltration; D017641:Zeolites / Q000737:chemistry* / Q000378:metabolism*
D011506:Proteins; D017641:Zeolites; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D006735:Horseradish Peroxidase; D009113:Muramidase