PEDS Advance Access originally published online on April 28, 2004
Protein Engineering Design and Selection 2004 17(4):293-304; doi:10.1093/protein/gzh038
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Directed evolution of an anti-carcinoembryonic antigen scFv with a 4-day monovalent dissociation half-time at 37°C
1Department of Chemical Engineering and Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, MA 02139, USA and 2Royal Free Hospital, University College London, Gower Street, London WC1E 6BT, UK
3 To whom correspondence should be addressed. E-mail: wittrup{at}mit.edu
An scFv has been engineered to bind carcinoembryonic antigen (CEA) with a dissociation half-time >4 days at 37°C. Two mutations responsible for this affinity increase were isolated by screening yeast surface-displayed mutant libraries by flow cytometry. Soluble expression of the mutant scFv in a yeast secretion system was increased 100-fold by screening mutant libraries for improved yeast surface display level. This scFv will be useful as a limiting case for evaluating the significance of affinity in tumor targeting to non-internalizing antigens.
Received December 17, 2003; revised April 2, 2004; accepted April 8, 2004.
Edited by Dario Neri
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