title: Antibodies against non-immunizing antigens derived from a large immune scFv library.
abstract: Target-specific antibodies can be rapidly enriched and identified from an antibody library using phage display. Large, naïve antibody libraries derived from synthetic or unimmunized sources can yield antibodies against virtually any antigen, whereas libraries from immunized sources tend to be smaller and are used exclusively against the antigen of immunization. In this study, 25 scFv libraries made from the spleens of immunized rabbits, each with a size ranging from 10(8) to higher than 10(9), were combined into a single large library with > 10(10) individual clones. Panning of this combined library yielded target-specific rabbit scFv clones against many non-immunizing antigens, including proteins, peptides, and a small molecule. Notably, specific scFv clones against a rabbit self-antigen (rabbit serum albumin) and a phosphorylated protein (epidermal growth factor receptor pTyr1173) could be isolated from the library. These results suggest that the immune library contained a significant number of unimmunized clones and that a sufficiently large immune library can be utilized similarly to a naïve library, i.e., against various non-immunizing antigens to yield specific antibodies.
authors.affiliation: Division of Life and Pharmaceutical Sciences, The Graduate School, Ewha Womans University, Seoul, 120-750, Korea.
authors.forename: Sung Ah
authors.initials: SA
authors.lastname: Moon
authors.forename: Min Kyung
authors.initials: MK
authors.lastname: Ki
authors.forename: Sungyoung
authors.initials: S
authors.lastname: Lee
authors.forename: Mi-Lim
authors.initials: ML
authors.lastname: Hong
authors.forename: Misook
authors.initials: M
authors.lastname: Kim
authors.forename: Sungsub
authors.initials: S
authors.lastname: Kim
authors.forename: Junho
authors.initials: J
authors.lastname: Chung
authors.forename: Sue Goo
authors.initials: SG
authors.lastname: Rhee
authors.forename: Hyunbo
authors.initials: H
authors.lastname: Shim
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000737:chemistry*; D002457:Cell Extracts / Q000737:chemistry; D066246:ErbB Receptors / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D019151:Peptide Library; D010750:Phosphoproteins / Q000737:chemistry; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000737:chemistry
chemical_list: D000941:Antigens; D002457:Cell Extracts; D019151:Peptide Library; D010750:Phosphoproteins; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D066246:ErbB Receptors