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Parallel barcoding of antibodies for DNA ‐assisted proteomics
Author(s) -
Dezfouli Mahya,
Vickovic Sanja,
Iglesias Maria Jesus,
Schwenk Jochen M.,
Ahmadian Afshin
Publication year - 2014
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201400215
Subject(s) - multiplex , proteomics , computational biology , proteome , dna , biology , dna sequencing , massive parallel sequencing , dna barcoding , microbiology and biotechnology , bioinformatics , gene , genetics , ecology
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.