Immobilization of perfluoroalkylated enzymes in a biologically active state onto Perflex support
Boivin, P.; Kobos, R.K.; Papa, S.L.; Scouten, W.H.
Biotechnology and Applied Biochemistry 14(2): 155-169
1991
ISSN/ISBN: 0885-4513 PMID: 1760128 Document Number: 378733
Perflex has been introduced by E.I. du Pont de Nemours and Co., Inc., as a new fluorocarbon-based technology for protein immobilization. Due to the hydrophobic character of the support, however, significant loss of enzymatic activity may occur upon immobilization of certain enzymes, which appears to be due to a large conformational change of the protein ("inversion"). Pretreatment of the Perflex support with a neutral fluorosurfactant lessened the surface hydrophobicity, thus decreasing the hydrophobic interaction between the support and the protein. Modification of enzymes with a high number of fluorocarbon residues, which forms a hydrophobic "envelope" around the protein, also appears to prevent enzyme inactivation upon immobilization on Perflex support. Moreover, preactivation of the support with either perfluorooctylpropylisocyanate or reactive poly(fluoroalkyl)sugar reagents greatly improves the enzyme particle activity by increasing the amount of immobilized enzyme. Fluorosurfactant treatment of the support activated with perfluorooctylpropylisocyanate improves the retention of activity for sensitive enzymes such as .alpha.-chymotrypsin and increases the wetability and ease of handling of the Perflex particles.