Arsenical grafted membranes for immobilization of thioredoxin-like proteins

• Published in: Reactive & functional polymers 2009-11, Vol.69 (11), p.816-820
• Main Authors: Carbajal, M. Laura, Espinoza, Silvia L. Soto, Valdez, Silvina N., Poskus, Edgardo, Smolko, Eduardo E., Grasselli, Mariano
• Format: Article
• Language: English
• Subjects: Applied sciences; Biological and medical sciences; Biotechnology; Exact sciences and technology; Exchange resins and membranes; Forms of application and semi-finished materials; Fundamental and applied biological sciences. Psychology; Immobilization of enzymes and other molecules; Immobilization techniques; Methods. Procedures. Technologies; Phenylarsenical ligands; Polymer industry, paints, wood; Protein immobilization; Radiation-induced graft polymerization; Technology of polymers; Thioredoxin; Thioredoxin-fusion protein
• ISSN: 1381-5148
• DOI: 10.1016/j.reactfunctpolym.2009.07.001

In this work, we describe the cografting of glycidyl methacrylate and dimethyl acrylamide onto a macroporous polysulfone polymer. Aminophenyl arsenical compounds were covalently attached to the copolymer through epoxy ring add-on reactions followed by a 2-mercaptoethanol activation. Thioredoxin and thioredoxin-fusion proteins were immobilized onto this surface and detected by specific antibody recognition. Preservation of native protein folding was confirmed by the detection of the enzymatic activity of an unstable fusion protein. Immobilized fusion protein onto the modified material maintains the enzymatic activity for a longer time, up to two weeks, against the free protein under the same storage conditions that remains active for 2 days.