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PEDS Advance Access published online on April 28, 2004
Protein Engineering Design and Selection, doi:10.1093/protein/gzh038
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Received December 17, 2003
Revised April 2, 2004
Accepted April 8, 2004

Article

Directed Evolution of an Anti-Carcinoembryonic Antigen scFv with a Four-Day Monovalent Dissociation Half-time at 37°C

Christilyn P. Graff 1, Kerry Chester 2, Richard Begent 2, K. Dane Wittrup 1*

1 Department of Chemical Engineering & Biological Engineering Division, Massachusetts Institute of Technology
2 Royal Free Hospital, University College London

* To whom correspondence should be addressed. E-mail: wittrup{at}mit.edu.


  Abstract

An scFv has been engineered to bind carcinoembryonic antigen (CEA) with a dissociation half-time over 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 one hundred-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 noninternalizing antigens.

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