A high-throughput pipeline for validation of antibodies. (November 2018)

• Record Type: Journal Article
• Authors: Sikorski, Krzysztof; Mehta, Adi; Inngjerdingen, Marit; Thakor, Flourina; Kling, Simon; Kalina, Tomas; Nyman, Tuula; Stensland, Maria; Zhou, Wei; Souza, Gustavo; Holden, Lars; Stuchly, Jan; Templin, Markus; Lund-Johansen, Fridtjof
• Is Part Of: Nature methods. Volume 15:Number 11(2018)
• Page Start: 909
• Page End: 912
• Publication Date: 2018-11
• Subjects: Life sciences -- Methodology -- Periodicals; Life sciences -- Research -- Periodicals; Biology -- Methodology -- Periodicals; Biology -- Research -- Periodicals; 570.72
• DOI: 10.1038/s41592-018-0179-8
• Languages: English
• ISSNs: 1548-7091

Abstract

Abstract Western blotting (WB) is widely used to test antibody specificity, but the assay has low throughput and precision. Here we used preparative gel electrophoresis to develop a capture format for WB. Fractions with soluble, size-separated proteins facilitated parallel readout with antibody arrays, shotgun mass spectrometry (MS) and immunoprecipitation followed by MS (IP-MS). This pipeline provided the means for large-scale implementation of antibody validation concepts proposed by an international working group on antibody validation (IWGAV). This paper describes a platform for carrying out antibody-based capture and mass spectrometry in parallel, and tests the feasibility of this platform for high-throughput validation of antibodies.