Antibodies against non-immunizing antigens derived from a large immune scFv library |
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Authors: | Sung Ah Moon Min Kyung Ki Sungyoung Lee Mi-Lim Hong Misook Kim Sungsub Kim Junho Chung Sue Goo Rhee Hyunbo Shim |
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Affiliation: | Division of Life and Pharmaceutical Sciences, The Graduate School, Ewha Womans University, Seoul, 120-750, Korea. |
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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 108 to higher than 109, were combined into a single large library with > 1010 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 naïe library, i.e., against various non-immunizing antigens to yield specific antibodies. |
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Keywords: | antigen specificity immune library non-immunizing antigen phage display scFv |
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