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 Helmsing; Michael 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

doi:10.1038/nmeth.1607

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

Research output: Contribution to journal › Article › Research › peer-review

273 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 language	English
Pages (from-to)	551 - 558
Number of pages	8
Journal	Nature Methods
Volume	8
Issue number	7
DOIs	https://doi.org/10.1038/nmeth.1607
Publication status	Published - 2011

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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

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title = "A roadmap to generate renewable protein binders to the human proteome",

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.",

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

year = "2011",

doi = "10.1038/nmeth.1607",

language = "English",

volume = "8",

pages = "551 -- 558",

journal = "Nature Methods",

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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, Helmsing, S, Hust, M, Dubel, S, Berkowicz, S, Freemantle, A, Spiegel, M, Sawyer, A, Layton, DS, Nice, E, Dai, A, Rocks, O, Williton, K, Fellouse, F, Hersi, K, Pawson, T, Nilsson, P, Sundberg, M, Sjoberg, R, Sivertsson, A, Schwenk, J, Takanen, J, Hober, S, Uhlen, M, Dahlgren, L-G, Flores, A, Johansson, I, Weigelt, J, Crombet, L, Loppnau, P, Kozieradzki, I, Cossar, D, Arrowsmith, CH, Edwards, A & Graslund, S 2011, 'A roadmap to generate renewable protein binders to the human proteome', Nature Methods, vol. 8, no. 7, pp. 551 - 558. https://doi.org/10.1038/nmeth.1607

TY - JOUR

T1 - A roadmap to generate renewable protein binders to the human proteome

AU - Colwill, Karen

AU - Persson, Helena

AU - Jarvik, Nicholas

AU - Wyrzucki, Arkadiusz

AU - Wojcik, John

AU - Koide, Akiko

AU - Kossiakoff, Anthony

AU - Koide, Shohei

AU - Sidhu, Sachdev

AU - Dyson, Michael

AU - Pershad, Kritika

AU - Pavlovic, John

AU - Karatt-Vellatt, Aneesh

AU - Schofield, Darren

AU - Kay, Brian

AU - McCafferty, John

AU - Mersmann, Michael

AU - Meier, Doris

AU - Mersmann, Jana

AU - Helmsing, Saskia

AU - Hust, Michael

AU - Dubel, Stefan

AU - Berkowicz, Susan

AU - Freemantle, Alexia

AU - Spiegel, Michael

AU - Sawyer, Alan

AU - Layton, Daniel Stuart

AU - Nice, Edouard

AU - Dai, Anna

AU - Rocks, Oliver

AU - Williton, Kelly

AU - Fellouse, Frederic

AU - Hersi, Kadija

AU - Pawson, Tony

AU - Nilsson, Peter

AU - Sundberg, Marten

AU - Sjoberg, Ronald

AU - Sivertsson, Asa

AU - Schwenk, Jochen

AU - Takanen, Jenny

AU - Hober, Sophia

AU - Uhlen, Mathias

AU - Dahlgren, Lars-Goran

AU - Flores, Alex

AU - Johansson, Ida

AU - Weigelt, Johan

AU - Crombet, Lissette

AU - Loppnau, Peter

AU - Kozieradzki, Ivona

AU - Cossar, Doug

AU - Arrowsmith, Cheryl H

AU - Edwards, Aled

AU - Graslund, Susanne

PY - 2011

Y1 - 2011

N2 - 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.

AB - 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.

UR - http://www.nature.com/nmeth/journal/v8/n7/pdf/nmeth.1607.pdf

U2 - 10.1038/nmeth.1607

DO - 10.1038/nmeth.1607

M3 - Article

SN - 1548-7091

VL - 8

SP - 551

EP - 558

JO - Nature Methods

JF - Nature Methods

IS - 7

ER -

A roadmap to generate renewable protein binders to the human proteome

Abstract

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