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Mining a yeast library for brain endothelial cell-binding antibodies

Nature Methods volume 4, pages 143–145 (2007)Cite this article

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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Figure 1: Identification of RBE4-binding scFv clones by cell panning and high-throughput scFv analysis.
Figure 2: Evaluation of scFv binding and internalization properties.

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References

  1. Popkov, M., Rader, C. & Barbas, C.F., III J. Immunol. Methods, 291, 137–151 (2004).

    Article  CAS  Google Scholar 

  2. Poul, M.A., Becerril, B., Nielsen, U.B., Morisson, P. & Marks, J.D. J. Mol. Biol. 301, 1149–1161 (2000).

    Article  CAS  Google Scholar 

  3. Ridgway, J.B. et al. Cancer Res. 59, 2718–2723 (1999).

    CAS  PubMed  Google Scholar 

  4. Valadon, P. et al. Proc. Natl. Acad. Sci. USA 103, 407–412 (2006).

    Article  CAS  Google Scholar 

  5. Muruganandam, A., Tanha, J., Narang, S. & Stanimirovic, D. FASEB J. 16, 240–242 (2002).

    Article  CAS  Google Scholar 

  6. Hoogenboom, H.R. Nat. Biotechnol. 23, 1105–1116 (2005).

    Article  CAS  Google Scholar 

  7. Wang, X.X. & Shusta, E.V. J. Immunol. Methods 304, 30–42 (2005).

    Article  CAS  Google Scholar 

  8. Feldhaus, M.J. et al. Nat. Biotechnol. 21, 163–170 (2003).

    Article  CAS  Google Scholar 

  9. Roux, F. et al. J. Cell. Physiol. 159, 101–113 (1994).

    Article  CAS  Google Scholar 

  10. Rakestraw, A. & Wittrup, K.D. Biotechnol. Bioeng. 93, 896–905 (2006).

    Article  CAS  Google Scholar 

  11. Hoogenboom, H.R. et al. Eur. J. Biochem. 260, 774–784 (1999).

    Article  CAS  Google Scholar 

  12. Shusta, E.V., Raines, R.T., Pluckthun, A. & Wittrup, K.D. Nat. Biotechnol. 16, 773–777 (1998).

    Article  CAS  Google Scholar 

  13. Friden, P.M. et al. Proc. Natl. Acad. Sci. USA 88, 4771–4775 (1991).

    Article  CAS  Google Scholar 

  14. Pardridge, W.M. NeuroRx 2, 3–14 (2005).

    Article  Google Scholar 

  15. Hackel, B.J., Huang, D., Bubolz, J.C., Wang, X.X. & Shusta, E.V. Pharm. Res. 23, 790–797 (2006).

    Article  CAS  Google Scholar 

Download references

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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Authors and Affiliations

  1. Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, Wisconsin, USA

    Xin Xiang Wang, Yong Ku Cho & Eric V Shusta

Authors
  1. Xin Xiang Wang
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  2. Yong Ku Cho
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  3. Eric V Shusta
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Contributions

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.

Corresponding author

Correspondence to Eric V Shusta.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Assessment of scFvA target antigen density and brain localization.

Supplementary Table 1

Summary of RBE4-binding scFv clones.

Supplementary Methods

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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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