Two stage screening of combinatorial libreries. Application to bovine serum

M. C. MARTÍNEZ CERON; S. L. GIUDICESSI; J. N. KRUZYN; M. M. MARANI; F. ALBERICIO; O. CASCONE; S. A. CAMPERI

Revista Cenic Ciencias Biológicas

CENIC

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

2221-2450

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 to Texas Red. Fluorescent beads were automatically isolated using the Complex Object Parametric Analyzer and Sorter (COPAS) equipment. Isolated beads were washed and the second screening was performed with BSA coupled to biotin and with streptavidin-peroxidase and revealed with 3,3´-diaminobenzidine. Peptides from isolated beads were sequenced using a mass spectrometry analyzer. Those peptides appearing with more frequency were synthesized and immobilized on agarose. All peptides adsorbed seroalbumin but not Texas-Red or streptavidin-peroxidase.