Journal
NATURE METHODS
Volume 8, Issue 7, Pages 551-U52Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.1607
Keywords
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Funding
- Canadian Institutes for Health Research
- Canadian Foundation for Innovation
- Genome Canada through the Ontario Genomics Institute
- GlaxoSmithKline
- Karolinska Institutet
- Knut and Alice Wallenberg Foundation
- Ontario Innovation Trust
- Ontario Ministry for Research and Innovation
- Merck Co., Inc.
- Novartis Research Foundation
- Swedish Agency for Innovation Systems
- Swedish Foundation for Strategic Research
- Wellcome Trust
- Ontario Research Fund Global Leadership Round in Genomics and Life Sciences
- Human Frontier Science Program
- European CommisSystematisch-Methodischen Platform Antibody Factory, within the German Nationales Genomforschungsnetzsion
- Land Niedersachsen
- US National Institutes of Health [GM082288-09A1, EY016094-01A1, R01-GM72688, U54-GM74946]
- Swedish Research Council [524-2008-617]
- Victorian State Government (The Department of Innovation, Industry and Regional Development)
- Australian Federal Government (Bioplatforms Australia)
- Monash University
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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.
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