title: Delineation of in vivo assembled multiprotein complexes via biomolecular interaction analysis mass spectrometry.
abstract: Biomolecular interaction analysis mass spectrometry (BIA-MS) is a multiplexed bioanalytical approach used in analysis of proteins from complex biological mixtures. It utilizes surface-immobilized ligands for protein affinity retrieval, surface plasmon resonance for monitoring the ligand-protein interaction and matrix-assisted laser desorption/ionization-time of flight mass spectrometry for revealing the masses of the biomolecules retrieved by the ligand. In order to explore the utility of BIA-MS in delineation of multiprotein complexes, an in vivo assembled protein complex comprised of retinol binding protein (RBP) and transthyretin (TTR) was investigated. Antibodies to RBP and TTR were utilized as ligands in the analysis of the protein complex present in human plasma. The RBP-TTR complex was retrieved by the anti-RBP antibody as indicated by the presence of both RBP and TTR signals in the mass spectra. RBP signals were not observed in the mass spectra of the material retained on the anti-TTR derivatized surface. In addition, the mass-specific detection in BIA-MS allowed detection of RBP and TTR analyte variants.
authors.affiliation: Intrinsic Bioprobes Inc, Tempe, AZ 85281, USA. dnedelkov@intrinsicbio.com
authors.forename: D
authors.initials: D
authors.lastname: Nedelkov
authors.forename: R W
authors.initials: RW
authors.lastname: Nelson
mesh_terms: D006801:Humans; D008024:Ligands; D008954:Models, Biological; D011228:Prealbumin / Q000737:chemistry; D011485:Protein Binding; D012177:Retinol-Binding Proteins / Q000737:chemistry; D054839:Retinol-Binding Proteins, Plasma; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D008024:Ligands; D011228:Prealbumin; D012177:Retinol-Binding Proteins; D054839:Retinol-Binding Proteins, Plasma