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A roadmap to generate renewable protein binders to the human proteome
KTH, Skolan för bioteknologi (BIO), Proteomik.ORCID-id: 0000-0002-4657-8532
KTH, Skolan för bioteknologi (BIO), Proteomik.
KTH, Skolan för bioteknologi (BIO), Proteomik.ORCID-id: 0000-0003-1363-5796
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Antal upphovsmän: 532011 (Engelska)Ingår i: Nature Methods, ISSN 1548-7091, E-ISSN 1548-7105, Vol. 8, nr 7, s. 551-8Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

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.

Ort, förlag, år, upplaga, sidor
Nature America Inc. , 2011. Vol. 8, nr 7, s. 551-8
Nyckelord [en]
Immunology, HIGH-THROUGHPUT GENERATION, PHAGE DISPLAY, RECOMBINANT ANTIBODIES, AFFINITY REAGENTS
Nationell ämneskategori
Naturvetenskap
Identifikatorer
URN: urn:nbn:se:kth:diva-74487DOI: 10.1038/nmeth.1607ISI: 000292194500016PubMedID: 21572409Scopus ID: 2-s2.0-79959851876OAI: oai:DiVA.org:kth-74487DiVA, id: diva2:489703
Forskningsfinansiär
Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Anmärkning
Author count: 53. QC 20120214Tillgänglig från: 2012-02-14 Skapad: 2012-02-03 Senast uppdaterad: 2017-12-08Bibliografiskt granskad

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Av författaren/redaktören
Nilsson, PeterSundberg, MårtenSjöberg, RonaldSivertsson, ÅsaSchwenk, Jochen MOttosson Takanen, JennyHober, SophiaUhlén, Mathias
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Proteomik
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Nature Methods
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