Two-stage screening of combinatorial peptide libraries. Application to bovine serum albumin ligand selection

MARTÍNEZ CERON MARÍA C.; GIUDICESSI SILVANA L.; KRUSZYN JOHANNA N.; MARANI MARIELA M.; ALBERICIO FERNANDO; CASCONE OSVALDO; CAMPERI SILVIA A.

Revista CENIC

Editorial CENIC

Lugar: La Habana; Año: 2015 vol. 46 p. 77 - 86

Short peptides are excellent ligands for affinity chromatrography. Divide-couple-recombine method allows obtaining a peptide library with all possible combinations of the amino acids in the form of "one bead-one peptide". In this work we designed a two-stage library screening method for peptide ligands selection. The first library screening was performed with the target protein (bovine seroalbumin) coupled with Texas Red. Fluorescent beads were automatically isolated using Complex Object Parametric Analyzer and Sorter (COPAS). Isolated beads were washed and the second screening was performed with BSA coupled with biotin and with streptavidin-peroxidase and revealed with 3,3´-diaminobenzidine. Those peptides that appear with more frequency were synthesized and immobilized on agarose. All peptide adsorbed seroalbumin but not adsorbed Texas-Red or streptavidin-peroxidase.