title: Parallel barcoding of antibodies for DNA-assisted proteomics.
abstract: 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.
authors.affiliation: Science for Life Laboratory, Division of Gene Technology, School of Biotechnology, Royal Institute of Technology (KTH), Solna, Sweden.
authors.forename: Mahya
authors.initials: M
authors.lastname: Dezfouli
authors.forename: Sanja
authors.initials: S
authors.lastname: Vickovic
authors.forename: Maria Jesus
authors.initials: MJ
authors.lastname: Iglesias
authors.forename: Jochen M
authors.initials: JM
authors.lastname: Schwenk
authors.forename: Afshin
authors.initials: A
authors.lastname: Ahmadian
mesh_terms: D000906:Antibodies / Q000737:chemistry*; D001798:Blood Proteins / Q000032:analysis*; D004247:DNA / Q000737:chemistry*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D007118:Immunoassay / Q000379:methods; D018796:Immunoconjugates / Q000737:chemistry; D018189:Immunomagnetic Separation / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D040901:Proteomics / Q000379:methods*
chemical_list: D000906:Antibodies; D001798:Blood Proteins; D018796:Immunoconjugates; D004247:DNA
keywords: Antibody; Bio-conjugation; DNA-assisted proteomics; DNA-barcoding; Massively parallel sequencing; Technology