title: Magnetic bead assisted labeling of antibodies at nanogram scale.
abstract: There are currently several initiatives that aim to produce binding reagents for proteome-wide analysis. To enable protein detection, visualization, and target quantification, covalent coupling of reporter molecules to antibodies is essential. However, current labeling protocols recommend considerable amount of antibodies, require antibody purity and are not designed for automation. Given that small amounts of antibodies are often sufficient for downstream analysis, we developed a labeling protocol that combines purification and modification of antibodies at submicrogram quantities. With the support of magnetic microspheres, automated labeling of antibodies in parallel using biotin or fluorescent dyes was achieved.
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: Peter
authors.initials: P
authors.lastname: Nilsson
authors.forename: Jochen M
authors.initials: JM
authors.lastname: Schwenk
authors.forename: Afshin
authors.initials: A
authors.lastname: Ahmadian
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry*; D001709:Biotechnology / Q000379:methods*; D001710:Biotin; D005456:Fluorescent Dyes; D018189:Immunomagnetic Separation / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*
chemical_list: D055910:Antibodies, Immobilized; D005456:Fluorescent Dyes; D001710:Biotin
keywords: Antibody; Automated protocol; Biotinylation; Labeling; Nanogram scale; Technology