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Yuan QA , Robinson MK , Simmons HH , Russeva M , Adams GP
Isolation of anti-MISIIR scFv molecules from a phage display library by cell sorter biopanning
Cancer Immunol Immunother. 2008 Mar;57(3) :367-78
PMID: 17676323   
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Abstract
While cell surface antigens represent the most common targets for antibody-based cancer therapy, isolation of new antibodies specific for these targets from single-chain Fv phage display libraries has been hindered by limitations associated with traditional selection techniques. Solid phase panning is often associated with conformational changes to the target protein due to its immobilization on plastic tubes that can limit the ability of the isolated scFv to bind to conformational epitopes and solution panning methods require the use of secondary tags that often mask desired sequences and create unintended epitopes. Commonly utilized cell-based panning methods typically yield a panel of single-chain Fv (scFv) molecules that are specific for numerous cell surface antigens, often obscuring the desired clones. Here, we describe a novel cell sorter-based system to isolate single-chain Fv molecules specific for defined antigen targets expressed on stably-transformed mammalian cells. We employed these methods to isolate promising scFv clones that bind specifically to the Mullerian inhibiting substance type II receptor, a cell surface ovarian cancer antigen that has proven to be a difficult target for selection strategies.
Notes
Yuan, Qing-An Robinson, Matthew K Simmons, Heidi H Russeva, Maria Adams, Gregory P P50 CA83638/CA/NCI NIH HHS/United States Research Support, N.I.H., Extramural Germany Cancer immunology, immunotherapy : CII Cancer Immunol Immunother. 2008 Mar;57(3):367-78. Epub 2007 Aug 4.