title: Simple method for production of randomized human tenth fibronectin domain III libraries for use in combinatorial screening procedures.
abstract: Challenges such as the rapid development of detection reagents for emerging or engineered pathogens, the goal of identifying probes for every protein in the human proteome, and the development of therapeutic molecules require systems for development of epitope binding molecules that are faster and cheaper than conventional antibody development. To be practical and effective, antibody mimics must be small, stable molecules that contain exposed loops or surfaces that can be randomized and screened using selective combinatorial assays. The tenth human fibronectin type III domain (10Fn3) fits these requirements and has recently been developed as an antibody mimic for use in detection and therapeutic platforms. Previously described systems for working with 10Fn3 used PCR-based approaches to anneal multiple oligonucleotides to generate randomized 10Fn3 libraries. Here we describe a simplified approach for creating randomized 10Fn3 libraries and report the first use of a T7-based phage display system for screening these libraries.
authors.affiliation: New Mexico State University, Las Cruces, NM 88003, USA.
authors.forename: David
authors.initials: D
authors.lastname: Garcia-Ibilcieta
authors.forename: Maxim
authors.initials: M
authors.lastname: Bokov
authors.forename: Vladimir
authors.initials: V
authors.lastname: Cherkasov
authors.forename: Petr
authors.initials: P
authors.lastname: Sveshnikov
authors.forename: Stephen F
authors.initials: SF
authors.lastname: Hanson
mesh_terms: D000595:Amino Acid Sequence; D017123:Bacteriophage T7 / Q000235:genetics; D020650:Combinatorial Chemistry Techniques; D005353:Fibronectins / Q000737:chemistry*; D006801:Humans; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019151:Peptide Library; D016133:Polymerase Chain Reaction
chemical_list: D005353:Fibronectins; D019151:Peptide Library