* To whom correspondence should be addressed.
Received June 5, 2008; Revision received September 10, 2008
State Research Center of Applied Microbiology and Biotechnology, Ministry of Health and Social Development of the Russian Federation, 142279 Obolensk, Moscow Region, Russia; fax (4967) 36-0010; E-mail: email@example.com
A method for adsorption chromatography of proteins is proposed. A protein solution is passed through a cellulose column at a pH value corresponding to an isoelectric point of the protein. Depending on the charge of unwanted proteins, they either remain at the origin (if charges of protein and ion-exchanger are opposite) or are released from the column (if charges of protein and ion-exchanger coincide). Elution volume of the purified protein is higher than for the second group of unwanted proteins because movement of the uncharged protein of interest includes its adsorption on cellulose followed by subsequent desorption caused by the elution buffer. Problems of optimization of buffers and adsorbents are discussed. Applicability of the method of adsorption chromatography is illustrated using purification of horseradish peroxidase as an example.
KEY WORDS: adsorption chromatography, adsorbent-ion exchanger, cellulose, buffer, peroxidase, amine