A roadmap to generate renewable protein binders to the human proteome

Karen Colwill, Helena Persson, Nicholas Jarvik, Arkadiusz Wyrzucki, John Wojcik, Akiko Koide, Anthony Kossiakoff, Shohei Koide, Sachdev Sidhu, Michael Dyson, Kritika Pershad, John Pavlovic, Aneesh Karatt-Vellatt, Darren Schofield, Brian Kay, John McCafferty, Michael Mersmann, Doris Meier, Jana Mersmann, Saskia HelmsingMichael Hust, Stefan Dubel, Susan Berkowicz, Alexia Freemantle, Michael Spiegel, Alan Sawyer, Daniel Stuart Layton, Edouard Nice, Anna Dai, Oliver Rocks, Kelly Williton, Frederic Fellouse, Kadija Hersi, Tony Pawson, Peter Nilsson, Marten Sundberg, Ronald Sjoberg, Asa Sivertsson, Jochen Schwenk, Jenny Takanen, Sophia Hober, Mathias Uhlen, Lars-Goran Dahlgren, Alex Flores, Ida Johansson, Johan Weigelt, Lissette Crombet, Peter Loppnau, Ivona Kozieradzki, Doug Cossar, Cheryl H Arrowsmith, Aled Edwards, Susanne Graslund

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133 Citations (Scopus)

Abstract

Despite the wealth of commercially available antibodies to human proteins, research is often hindered by their inconsistent validation, their poor performance and the inadequate coverage of the proteome. These issues could be addressed by systematic, genome-wide efforts to generate and validate renewable protein binders. We report a multicenter study to assess the potential of hybridoma and phage-display technologies in a coordinated large-scale antibody generation and validation effort. We produced over 1,000 antibodies targeting 20 SH2 domain proteins and evaluated them for potency and specificity by enzyme-linked immunosorbent assay (ELISA), protein microarray and surface plasmon resonance (SPR). We also tested selected antibodies in immunoprecipitation, immunoblotting and immunofluorescence assays. Our results show that high-affinity, high-specificity renewable antibodies generated by different technologies can be produced quickly and efficiently. We believe that this work serves as a foundation and template for future larger-scale studies to create renewable protein binders.
Original languageEnglish
Pages (from-to)551 - 558
Number of pages8
JournalNature Methods
Volume8
Issue number7
DOIs
Publication statusPublished - 2011

Cite this

Colwill, K., Persson, H., Jarvik, N., Wyrzucki, A., Wojcik, J., Koide, A., Kossiakoff, A., Koide, S., Sidhu, S., Dyson, M., Pershad, K., Pavlovic, J., Karatt-Vellatt, A., Schofield, D., Kay, B., McCafferty, J., Mersmann, M., Meier, D., Mersmann, J., ... Graslund, S. (2011). A roadmap to generate renewable protein binders to the human proteome. Nature Methods, 8(7), 551 - 558. https://doi.org/10.1038/nmeth.1607