6533b85ffe1ef96bd12c1389

RESEARCH PRODUCT

Revisited BIA-MS combination: Entire "on-a-chip" processing leading to the proteins identification at low femtomole to sub-femtomole levels

Géraldine LucchiPatrick DucoroyAlain RouleauWilfrid Boireau

subject

In situMALDI-TOFAnalyte[ SDV.BBM.BP ] Life Sciences [q-bio]/Biochemistry Molecular Biology/BiophysicsBiomedical EngineeringBiophysicsAnalytical chemistrySPRBiosensing TechniquesMass spectrometry01 natural sciencesSensitivity and Specificity03 medical and health sciencesProtein Interaction MappingElectrochemistryNanotechnologyBIA-MSBiochipChromatography High Pressure Liquid030304 developmental biology0303 health sciencesChromatographyprotein complexesElutionChemistryMicrochemistry010401 analytical chemistryMs analysisReproducibility of ResultsGeneral MedicineEquipment DesignMicrofluidic Analytical Techniques0104 chemical sciencesEquipment Failure Analysis[SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry Molecular Biology/BiophysicsMatrix-assisted laser desorption/ionizationSAMSpectrometry Mass Matrix-Assisted Laser Desorption-IonizationBiotechnology

description

International audience; We present the results of a study in which biomolecular interaction analysis (BIA, Biacore 2000) was combined with mass spectrometry (MS) using entire "on-a-chip" procedure. Most BIA-MS studies included an elution step of the analyte prior MS analysis. Here, we report a low-cost approach combining Biacore analysis with homemade chips and MS in situ identification onto the chips without elution step. First experiments have been made with rat serum albumin to determine the sensitivity and validation of the concept has been obtained with an antibody/antigen couple. Our "on-a-chip" procedure allowed complete analysis by MS-MS of the biochip leading to protein identifications at low femtomole to sub-femtomole levels. Using this technique, identification of protein complexes were routinely obtained giving the opportunity to the "on-a-chip" processing to complete the BIA-MS approach in the discovery and analysis of protein complexes

10.1016/j.bios.2008.06.030https://hal.archives-ouvertes.fr/hal-00347652/document