title: Bovine serum albumin as a universal suppressor of non-specific peptide binding in vials prior to nano-chromatography coupled mass-spectrometry analysis.
abstract: Non-specific binding (NSB) is a well-known problem for any application that deals with ultralow analyte quantities. The modern nano-flow chromatography coupled tandem mass-spectrometry (nanoLC-MS/MS) works with the lowest conceivable analyte concentrations. However, while the NSB problem is widely accepted and investigated for metabolomics and single-peptide medicine-related assays, its impact is not studied for complex peptide mixtures in proteomic applications. In this work peptide NSB to a common plastic autosampler vial was studied for a model mixture of 46 synthetic peptides. A significant NSB level was demonstrated for total peptide concentrations of up to 1 mg mL(-1). Different agents were tried for NSB suppression and compatibility with nanoLC-MS/MS analysis: a chaotropic agent, an amino acid mixture, a peptide mixture and a protein solution. The first two were inefficacious. The peptide matrix blocked NSB, however, it also led to analyte ionization suppression in nanoLC-MS/MS. The protein solution (0.1% BSA) efficiently eliminated NSB, while a trap-elute nanoHPLC configuration together with a small-pore reverse-phased sorbent effectively and quantitatively extracted the model peptides and depleted protein material from the sample. Higher protein concentration partially impeded peptide extraction. Thus, the 0.1% BSA solution might be regarded as an effective non-interfering blockader of NSB for sample resuspension and storage in an autosampler prior to LC-MS/MS analysis.
authors.affiliation: Laboratory of Proteomics, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 16/10 Miklukho-Maklaya str., Moscow 117997, Russian Federation; Laboratory of Proteomic Analysis, Research Institute of Physical Chemical Medicine, 1a Malaya Pirogovskaya str., Moscow 119435, Russian Federation. Electronic address: xerx222@gmail.com.
authors.forename: Sergey I
authors.initials: SI
authors.lastname: Kovalchuk
authors.affiliation: Laboratory of Proteomics, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 16/10 Miklukho-Maklaya str., Moscow 117997, Russian Federation.
authors.forename: Nikolay A
authors.initials: NA
authors.lastname: Anikanov
authors.affiliation: Laboratory of Proteomics, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 16/10 Miklukho-Maklaya str., Moscow 117997, Russian Federation.
authors.forename: Olga M
authors.initials: OM
authors.lastname: Ivanova
authors.affiliation: Laboratory of Proteomics, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 16/10 Miklukho-Maklaya str., Moscow 117997, Russian Federation.
authors.forename: Rustam H
authors.initials: RH
authors.lastname: Ziganshin
authors.affiliation: Laboratory of Proteomics, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 16/10 Miklukho-Maklaya str., Moscow 117997, Russian Federation; Laboratory of Proteomic Analysis, Research Institute of Physical Chemical Medicine, 1a Malaya Pirogovskaya str., Moscow 119435, Russian Federation.
authors.forename: Vadim M
authors.initials: VM
authors.lastname: Govorun
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003902:Detergents / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D036103:Nanotechnology; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D040901:Proteomics; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D000596:Amino Acids; D003902:Detergents; D010455:Peptides; D012710:Serum Albumin, Bovine
keywords: Nano chromatography coupled tandem mass-spectrometry; Non-specific binding; Peptidomics; Proteomics