Parallel barcoding of antibodies for DNA-assisted proteomics
2014 (English)In: Proteomics, ISSN 1615-9853, E-ISSN 1615-9861, Vol. 14, no 21-22, 2432-2436 p.Article in journal (Refereed) Published
DNA-assisted proteomics technologies enable ultra-sensitive measurements in multiplex format using DNA-barcoded affinity reagents. Although numerous antibodies are available, nowadays targeting nearly the complete human proteome, the majority is not accessible at the quantity, concentration, or purity recommended for most bio-conjugation protocols. Here, we introduce a magnetic bead-assisted DNA-barcoding approach, applicable for several antibodies in parallel, as well as reducing required reagents quantities up to a thousand-fold. The success of DNA-barcoding and retained functionality of antibodies were demonstrated in sandwich immunoassays and standard quantitative Immuno-PCR assays. Specific DNA-barcoding of antibodies for multiplex applications was presented on suspension bead arrays with read-out on a massively parallel sequencing platform in a procedure denoted Immuno-Sequencing. Conclusively, human plasma samples were analyzed to indicate the functionality of barcoded antibodies in intended proteomics applications.
Place, publisher, year, edition, pages
2014. Vol. 14, no 21-22, 2432-2436 p.
Antibody, Bio-conjugation, DNA-assisted proteomics, DNA-barcoding, Massively parallel sequencing, Technology
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-157615DOI: 10.1002/pmic.201400215ISI: 000344807600007PubMedID: 25263329ScopusID: 2-s2.0-84909579700OAI: oai:DiVA.org:kth-157615DiVA: diva2:771097
FunderVinnovaKnut and Alice Wallenberg FoundationScience for Life Laboratory - a national resource center for high-throughput molecular bioscience
QC 201412122014-12-122014-12-112015-02-03Bibliographically approved