Abstract
We describe the use of yeast surface display for the identification of antibodies that bind the plasma membranes of living cells. Yeast panning with a nonimmune human single-chain antibody library identified 34 unique lead antibodies that bind (Kd = 82 ± 15 nM) and in some cases internalize into rat brain endothelial cells. In addition, we used a new yeast display immunoprecipitation procedure for initial characterization of the cognate antigens.
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Acknowledgements
This work was supported by grants from the Whitaker Foundation (RG-02-0077), the Camille and Henry Dreyfus Foundation, and the US National Institutes of Health (NS052649). Y.K.C. was supported in part by the Samsung Lee Kun Hee Scholarship Foundation.
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X.X.W. and Y.K.C. performed the experiments. X.X.W., Y.K.C. and E.V.S designed the experiments and wrote the paper.
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Supplementary information
Supplementary Fig. 1
Assessment of scFvA target antigen density and brain localization.
Supplementary Table 1
Summary of RBE4-binding scFv clones.
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Wang, X., Cho, Y. & Shusta, E. Mining a yeast library for brain endothelial cell-binding antibodies. Nat Methods 4, 143–145 (2007). https://doi.org/10.1038/nmeth993
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DOI: https://doi.org/10.1038/nmeth993
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