Use of dextrans as long and hydrophilic spacer arms to improve the performance of immobilized proteins acting on macromolecules.
New dextran-agarose supports, suitable for covalent immobilization of enzymes and proteins acting on macromolecular substrates, were prepared. The thick internal fibers of agarose gels were covered by a low-density layer of long, flexible, hydrophilic, and inert dextran molecules. Rennin and protein A were immobilized on these novel supports and the resulting derivatives exhibited a very high capacity for biological recognition of soluble macromolecular substrates. Caseinolytic activity of this immobilized enzyme was 15-fold higher than activity of directly immobilized rennin, through short spacer arms, on agarose gels. Similarly, the new derivatives of immobilized protein A were able to adsorb up to 2 molecules of immunoglobulin per each molecule of immobilized protein A. When the immobilized proteins were secluded away from the support surface by using these new long and hydrophilic spacer arms, they exhibit minimal steric hindrances that could be promoted by the proximity of the support surface.
Penzol
G
G
Departamento de Biocatálisis, Instituto de Catálisis, CSIC, 28049 Madrid, Spain.
Armisén
P
P
Fernández-Lafuente
R
R
Rodés
L
L
Guisán
J M
JM
D000327:Adsorption; D000447:Aldehydes; D002364:Caseins / Q000378:metabolism; D012085:Chymosin / Q000378:metabolism; D003911:Dextrans; D004800:Enzymes, Immobilized / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D007136:Immunoglobulins; D012685:Sepharose; D013205:Staphylococcal Protein A
D000447:Aldehydes; D002364:Caseins; D003911:Dextrans; D004800:Enzymes, Immobilized; D007136:Immunoglobulins; D013205:Staphylococcal Protein A; D012685:Sepharose; D012085:Chymosin