title: Hyphenation of surface plasmon resonance imaging to matrix-assisted laser desorption ionization mass spectrometry by on-chip mass spectrometry and tandem mass spectrometry analysis.
abstract: Most of the recent developments aiming to the coupling between surface plasmon resonance (SPR) and mass spectrometry (MS) are based on the use of a biochip with a limited number of flow cells requiring elution steps for the recovery of the captured biomolecules. In this work, a direct on-chip MALDI-MS detection is presented using a SPRi-sensor biochip in a microarray format that allows a multiplex SPR-MS analysis. The biochip gold surface was functionalized by a self-assembled monolayer (SAM) of short polyoxyethylene (POE) chains carrying a N-hydroxysuccinimide (NHS) group for the immobilization of biomolecules. The SPR measurement of the interaction of grafted antibodies anti-beta-lactoglobulin and anti-ovalbumin with their corresponding antigens indicated that the POE-NHS SAM preserved the binding activity of the antibodies immobilized on the biochips surface. SPR-MS experiments were carried out through MALDI-MS detection of the retained antigens (beta-lactoglobulin and ovalbumin) directly from the biochip surface. Mass spectra were obtained from each distinct spot on the arrayed biochips. Femtomole amounts of specifically retained antigen proteins as determined by SPR were sufficient to obtain good quality mass spectra. These mass spectra showed protein ions corresponding to the specific antigen, without any trace of nonspecific binding. The underivatized portion of the chip was also devoid of nonspecifically bound proteins, indicating that the functionalization of the biochips surface by short polyoxyethylene chains greatly minimizes the unspecific binding. In addition, it allowed on-chip digestion of the specifically bound analyte and coupling with MS/MS experiments, opening numerous applications in the proteomic field.
authors.affiliation: CNRS, UMR8587, Université d'Evry-Val-d'Essonne, Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement, 91025 Evry, France.
authors.forename: S
authors.initials: S
authors.lastname: Bellon
authors.forename: W
authors.initials: W
authors.lastname: Buchmann
authors.forename: F
authors.initials: F
authors.lastname: Gonnet
authors.forename: N
authors.initials: N
authors.lastname: Jarroux
authors.forename: M
authors.initials: M
authors.lastname: Anger-Leroy
authors.forename: F
authors.initials: F
authors.lastname: Guillonneau
authors.forename: R
authors.initials: R
authors.lastname: Daniel
mesh_terms: D000595:Amino Acid Sequence; D055910:Antibodies, Immobilized / Q000276:immunology / Q000378:metabolism; D007782:Lactoglobulins / Q000032:analysis*; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation* / Q000379:methods; D013388:Succinimides / Q000737:chemistry; D020349:Surface Plasmon Resonance / Q000379:methods*; D013499:Surface Properties; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D055910:Antibodies, Immobilized; D007782:Lactoglobulins; D013388:Succinimides; D010047:Ovalbumin; C001426:N-hydroxysuccinimide