CAMPERI Silvia Andrea
congresos y reuniones científicas
Immobilized peptide as affinity ligand. Soybean peroxidase purification by affinity chromatography.
NANCY B. IANNUCCI; SILVIA A. CAMPERI; MARÍA V. MIRANDA; FERNANDO ALBERICIO; OSVALDO CASCONE
Simposio; 22nd American Peptide Society Symposium; 2011
Americam Peptide Society
An affinity peptide ligand for the soybean peroxidase (SBP) was identified from the screening of a combinatorial library composed by 130.321 tetrapeptides. All natural amino acids except cysteine were the library monomers and PEGA resin was the support. The selected peptide -KVQN- was synthesized on CLEARTM resin and the chromatographic parameters were calculated from the isotherm, resulting a Kd of 3x10-7 M and a maximum capacity of 12 mg SBP/ml resin. A cysteine residue was added to the C-terminal of the tetrapeptide with the aim of immobilize it on an agarose support by a thioether bond (SulphoLink SepharoseTM). This strategy optimized the high salt elution step, increasing the yield of the SBP recovery. Specificity assays were carried out. Alternative applications for this immobilized peptide are analyzed regarding the developing color feature of SBP.