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Multiplexed antigen bead arrays for the assessment of antibody selectivity and epitope mapping
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Affinity Proteomics. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0001-7843-2960
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Affinity Proteomics. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0002-4657-8532
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Affinity Proteomics. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0001-8141-8449
2018 (English)In: Epitope Mapping Protocols, Humana Press Inc. , 2018, p. 239-248Chapter in book (Refereed)
Abstract [en]

With the increasing number of binding reagents for affinity-based investigations of the human proteome, high-throughput tools for the characterization of the used reagents become essential. For the analysis of binding selectivity, bead-based antigen arrays offer a miniaturized and parallelized assay platform to meet such needs, as they enable two-dimensional multiplexing to analyze up to 384 samples against up to 500 analytes in a single round of analysis. In this chapter, we describe our protocols for the generation of multiplex bead arrays built on immobilized protein fragments, as well as biotinylated peptides. Combined together, these two versions of antigen arrays offer a versatile approach for multiplexed characterization of antibody binding selectivity, off-target interactions, as well as mapping for the amino acids of epitopes involved in antibody binding.

Place, publisher, year, edition, pages
Humana Press Inc. , 2018. p. 239-248
Series
Methods in Molecular Biology, ISSN 1064-3745 ; 1785
Keywords [en]
Affinity proteomics, Antibody selectivity, Antigen arrays, Epitope mapping, Peptide arrays, Protein arrays, Suspension bead arrays
National Category
Other Industrial Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-238377DOI: 10.1007/978-1-4939-7841-0_16Scopus ID: 2-s2.0-85046341154OAI: oai:DiVA.org:kth-238377DiVA, id: diva2:1264882
Funder
Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Note

QC 20181121

Available from: 2018-11-21 Created: 2018-11-21 Last updated: 2018-11-21Bibliographically approved

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