Inmovilización superficial de proteínas usando irradiación gamma

CARBAJAL, M.L.; HIDALGO, R.; BRITOS, C.; SMOLKO, E.E.; GRASSELLI, M.

Bs. As. Argentina

Congreso; XXXI Reunión Anual de la Asociación Argentina de Tecnología Nuclear (AATN).; 2004

Asociación Argentina de Tecnología Nuclear (AATN)

The modification of polymers by radiation grafting has been already reported for severaldecades. Nevertheless, the facility to tailoring surfaces to immobilize proteins remainsas an area of potential interest.In this work, we described the grafting of glycidyl methacrylate (GMA) and dimethtylacrylamide (DMAAm) onto a polysulfone membrane. Amount of grafting, from 10 to60% was obtained by choosing an initially favorable monomer concentration andgamma radiation doses of the order of 5 kGy. The addition of DMAAm to the monomersolution yields a new grafted copolymer with enhanced hydrophilic properties. Thepresence of grafted polymers were analyzed by FT-IR ATR spectroscopy.The subsequent immobilization of p-[bis-(β-hydroxyethylmercapto)-arsino]-aniline(PBMA) through a covalent attached to the epoxy moiety of grafted polymers wasperformed. Later on, recombinant thioredoxin protein (rTrx) and a Trx-fusion proteinwere immobilized onto this surface by a co-ordination site between PBMA and twoproximal cysteines belong to the protein structures. The presence of the proteins on thepolymer were detected by the attachment of specific antibodies.