Affinity polymeric support to thioredoxin. New materials for biotechnology applications

CARBAJAL, M.L.; VALDEZ, S.N.; VILLANUEVA, A.L.; POSKUS, E.; SMOLKO, E.E; GRASSELLI, M.

Villa Carlos Paz, Córdoba, Argentina.

Congreso; XXXVII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular (SAIBBM).; 2001

Sociedad Argentina de Investigación Bioquímica y Biología Molecular (SAIBBM).

Advanced functional materials are in high demand for a variety of applications ranging from catalysis to chromatography, diagnostics and sensors. Here we report the synthesis of a novel polymeric support with affinity to Thioredoxin (Trx). A polymeric membrane was coated with a copolymer based on glycidylmethacrylate and dimethylacrylamide. Onto its surface a specific Trx ligand (4-aminobenzenearsonic acid) was immobilised in two reaction steps. A minimum quantity of 0.5 ng Trx was detected on 1 cm2 of affinity membrane by a colorimetric detection procedure using an anti-Trx-antibody followed by a second peroxidase-conjugated-antibody.On the other hand, a Glutamic Acid Decarboxylase 65-Thioredoxin (Trx-GAD65) chimera was previously developed by our group as an effective antigen for GAD antibody (GADA) detection. GADA is an early marker for diagnostic support of insulin-dependent (Type 1) diabetes mellitus*. This fusion protein must be in the appropriate conformational state for its recognition by GADA, mainly directed to discontinuous conformational epitopes.*M.L: Papouchado, S.N. Valdez, M.R. Ermácora, S. Gañan and E. Poskus. J. Immunol. Meth. 207 (1997) 169-178.