Molecularly imprinted poly β-cyclodextrin polymer: Application in protein refolding

• Published in: Biochimica et biophysica acta Vol. 1770; no. 6; pp. 943 - 950
• Main Authors: Ali Esmaeili, Mohammad, Yazdanparast, Razieh
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.06.2007
• Subjects: Alkaline phosphatase; Alkaline Phosphatase - chemistry; beta-Cyclodextrins - chemistry; CHAPS; Cholic Acids - chemistry; Detergents - chemistry; Molecularly imprinted poly (β-CD) polymer; Protein Folding; Refolding Kinetic; β-cyclodextrin; Alkaline phosphatase; ALP; ANS; pNPP; CTAB; β-cyclodextrin; CHAPS; β-CD; THF; MIP; TDI; Refolding Kinetic; Molecularly imprinted poly (β-CD) polymer; GuHCl
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2007.02.007

Regarding our previous report on refolding of alkaline phosphatase [Yazdanparast and Khodagholi, 2005 Arch. Biochem. Biophys] it was found that in spite of the anti-aggregatory effect of 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), a zwitteronic detergent, the recovered activity was almost the same as the recovered activity obtained through the unassisted approach. The low recovery yield is probably due to the bulky groups of the detergent that interfere with its entrance into the small cavity of the stripping agent, cyclodextrin, implying that the stripping of detergent molecules from the detergent–protein complexes plays a major role in successful refolding processes. To improve the efficiency of CHAPS stripping, we evaluated, for the first time, the stripping potential of a molecular imprinting polymer designed to replace β-CD. In this approach, CHAPS was used as the template and the refolding of GuHCl denatured alkaline phosphatase was studied. Our results indicated that under the optimally developed refolding environment and similar to stripping by soluble β-CD, a refolding yield of 79% was obtained for denatured alkaline phosphatase using 20 mg/ml of the molecularly imprinted poly (β-CD) polymer. The major advantage of the new stripping agent, besides of its recycling option and ease of separation from the finished product, is its high potential of preventing aggregate formation. Based on these results, it seems that the new stripping strategy can constitute an ideal approach for refolding of proteins at much lower industrial costs compared to stripping with soluble β-cyclodextrin.