The Bead blot: a method for identifying ligand-protein and protein-protein interactions using combinatorial libraries of peptide ligands.

摘要: Small molecules that bind proteins can be used as ligands for protein purification and investigating protein-protein protein-drug interactions. Unfortunately, many methods to identify new desired suffer from common shortcomings, including the requirement target purified and/or selected under conditions different those which it will used. We have developed a method called Bead blot (i) select unpurified proteins, trace present in complex materials (e.g., unfractionated plasma); (ii) multiple variety of single experiment; (iii) with libraries types ligands. In blot, library ligands, synthesized on chromatography resin beads, is incubated starting material containing ligand sought. The their complementary according specific affinity Then protein-loaded beads are immobilized porous matrix, directionally eluted captured membrane superimposed beads. location determined, because position protein(s) reflects bead(s) bead originally bound identified, subsequent elucidation sequence. Ligands several targets identified one experiment. Here we demonstrate broad utility this by selection purify plasma complexes or remove pathogens whole blood very high constants. also based competitive elution.