10.1016/j.bmc.2008.09.061
The research focuses on developing an efficient one-cycle affinity selection method using a T7 phage display pool to identify binding proteins or peptides specific for small molecules. The study employs a cuvette type 27-MHz quartz-crystal microbalance (QCM) apparatus with a self-assembled monolayer (SAM) for immobilizing specific small molecules on the gold electrode surface of a sensor chip. The researchers used this system to screen for proteins or peptides that bind to synthetic ligand for FK506-binding protein (SLF) or the anti-tumor drug irinotecan (Iri, CPT-11). They successfully detected FK506-binding protein 12 (FKBP12)-displaying T7 phage with a short interaction time of 10 minutes, avoiding extensive wash or elution conditions and multiple rounds of selection. Additionally, a random-peptide T7 phage pool was utilized along with RELIC software for bioinformatics analysis, which helped pinpoint specific amino acid residues within the binding site of FKBP12 and regions of the Iri-binding site on known targets, acetylcholinesterase (AChE), and carboxylesterase (CE). The study demonstrates the method's effectiveness and broad applicability for screening small-molecule binding proteins/peptides. Reactants included SLF and Iri derivatives synthesized for forming SAM, natural-protein and random-peptide T7 phage pools, and His-tagged FKBP12. Analyses involved QCM monitoring, agarose gel electrophoresis, DNA sequencing, and bioinformatics tools like RELIC software.
