Enhanced validation of antibodies for research applications

Fredrik Edfors; Andreas Hober; Klas Linderbäk; Gianluca Maddalo; Alireza Azimi; Åsa Sivertsson; Hanna Tegel; Sophia Hober; Cristina Al Khalili Szigyarto; Linn Fagerberg; Kalle von Feilitzen; Per Oksvold; Cecilia Lindskog; Björn Forsström; Mathias Uhlén

doi:10.1038/s41467-018-06642-y

Enhanced validation of antibodies for research applications

Fredrik Edfors, Andreas Hober, Klas Linderbäk, Gianluca Maddalo, Alireza Azimi, Åsa Sivertsson, Hanna Tegel, Sophia Hober, Cristina Al Khalili Szigyarto, Linn Fagerberg, Kalle von Feilitzen, Per Oksvold, Cecilia Lindskog, Björn Forsström, Mathias Uhlén^*

^*Corresponding author for this work

Novo Nordisk Foundation Center for Biosustainability

Research output: Contribution to journal › Journal article › Research › peer-review

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Abstract

There is a need for standardized validation methods for antibody specificity and selectivity. Recently, five alternative validation pillars were proposed to explore the specificity of research antibodies using methods with no need for prior knowledge about the protein target. Here, we show that these principles can be used in a streamlined manner for enhanced validation of research antibodies in Western blot applications. More than 6,000 antibodies were validated with at least one of these strategies involving orthogonal methods, genetic knockdown, recombinant expression, independent antibodies, and capture mass spectrometry analysis. The results show a path forward for efforts to validate antibodies in an application-specific manner suitable for both providers and users.

Original language	English
Article number	4130
Journal	Nature Communications
Volume	9
ISSN	2041-1723
DOIs	https://doi.org/10.1038/s41467-018-06642-y
Publication status	Published - 2018

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Open Access This article is licensed under a Creative Commons Attribution 4.0 International License

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AU - Hober, Andreas

AU - Linderbäk, Klas

AU - Maddalo, Gianluca

AU - Azimi, Alireza

AU - Sivertsson, Åsa

AU - Tegel, Hanna

AU - Hober, Sophia

AU - Szigyarto, Cristina Al Khalili

AU - Fagerberg, Linn

AU - von Feilitzen, Kalle

AU - Oksvold, Per

AU - Lindskog, Cecilia

AU - Forsström, Björn

AU - Uhlén, Mathias

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N2 - There is a need for standardized validation methods for antibody specificity and selectivity. Recently, five alternative validation pillars were proposed to explore the specificity of research antibodies using methods with no need for prior knowledge about the protein target. Here, we show that these principles can be used in a streamlined manner for enhanced validation of research antibodies in Western blot applications. More than 6,000 antibodies were validated with at least one of these strategies involving orthogonal methods, genetic knockdown, recombinant expression, independent antibodies, and capture mass spectrometry analysis. The results show a path forward for efforts to validate antibodies in an application-specific manner suitable for both providers and users.

AB - There is a need for standardized validation methods for antibody specificity and selectivity. Recently, five alternative validation pillars were proposed to explore the specificity of research antibodies using methods with no need for prior knowledge about the protein target. Here, we show that these principles can be used in a streamlined manner for enhanced validation of research antibodies in Western blot applications. More than 6,000 antibodies were validated with at least one of these strategies involving orthogonal methods, genetic knockdown, recombinant expression, independent antibodies, and capture mass spectrometry analysis. The results show a path forward for efforts to validate antibodies in an application-specific manner suitable for both providers and users.

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DO - 10.1038/s41467-018-06642-y

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Enhanced validation of antibodies for research applications

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