Development of an on-line SPR-digestion-nanoLC-MS/MS system for the quantification and identification of interferon-γ in plasma

Stigter, E.C.A.; Jong, G.J. de; Bennekom, W.P. van

doi:10.1016/j.bios.2008.11.020

Cited by 22 publications

(12 citation statements)

References 27 publications

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“…In contrast, SPR-LC-MS (MS/MS) combinations have been previously reported, in which either proteolytic peptides of proteins isolated on an SPR chip were analyzed, or captured molecules reanalyzed by LC-MS [15][16][17]. Offline isolation of biosensor detected ligands followed by MALDI-MS has also been reported, but the relation to the biosensor signal quantification is lost in this approach [18,19].…”

Section: Interface For On-line Bioaffinity-esi-msmentioning

confidence: 99%

On-line bioaffinity-electrospray mass spectrometry for simultaneous detection, identification, and quantification of protein-ligand interactions

Drăguşanu

Petre

Slamnoiu

et al. 2010

J. Am. Soc. Mass Spectrom.

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We describe here an on-line combination of a surface acoustic wave (SAW) biosensor with electrospray ionization mass spectrometry (SAW-ESI-MS) that enables the direct detection, identification, and quantification of affinity-bound ligands from a protein-ligand complex on a biosensor chip. A trapping column was used between the SAW-biosensor and the electrospray mass spectrometer equipped with a micro-guard column, which provides simultaneous sample concentration and desalting for the mass spectrometric analysis of the dissociated ligand. First applications of the on-line SAW-ESI-MS combination include (1), differentiation of ␤-amyloid (A␤) epitope peptides bound to anti-A␤ antibodies; (2), the identification of immobilized Substance P peptide-calmodulin complex; (3), identification and quantification of the interaction of 3-nitrotyrosine-modified peptides with nitrotyrosine-specific antibodies; and (4), identification of immobilized anti-␣-synuclein-human ␣-synuclein complex. Quantitative determinations of protein-ligand complexes by SAW yielded dissociation constants (K D ) from micro-to low nanomolar sample concentrations. The on-line bioaffinity-ESI-MS combination presented here is expected to enable broad bioanalytical application to the simultaneous, label-free determination and quantification of biopolymer-ligand interactions, as diverse as antigen-antibody and lectin-carbohydrate complexes. (J Am Soc Mass Spectrom 2010, 21, 1643-1648 [3,4]. In particular, SPR has developed into an efficient tool for analysis of biomolecular recognition processes at a biosensor surface, and has been applied to the quantification of a variety of biopolymer interactions [4,5]. A recently-explored alternative to SPR is the surface acoustic wave (SAW) technology in which the piezoelectric effect of mass differences is employed for bioaffinity detection [6 -9]. SAW is now becoming increasingly important for the study of biomacromolecular interactions due to its high detection sensitivity in dilute solutions.Advantages of SAW in comparison to classical immuno-analytical techniques are the direct and rapid determination of association/dissociation constants with small sample amounts, and without labeling approaches or recalibration for buffer changes being required [6]. While providing sensitive and accurate determinations of binding/dissociation constants (K i or K D ), a major limitation of all bioaffinity methods is the lack of direct identification of the affinity-bound ligands. In contrast, the combination of biosensor detection and mass spectrometry enables both identification and quantification of bioaffinity interactions of biopolymers. Here we describe an on-line combination of an SAW biosensor with electrospray ionization mass spectrometry, SAW-ESI-MS. The on-line coupling between the SAW-sensor chip and the ESI-MS source was achieved by a incorporating a standard trapping column setup, using a six-port valve, a guard column, and a micropump system. The six-port valve interface provides both ligand concentration and an...

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Section: Interface For On-line Bioaffinity-esi-msmentioning

confidence: 99%

On-line bioaffinity-electrospray mass spectrometry for simultaneous detection, identification, and quantification of protein-ligand interactions

Drăguşanu

Petre

Slamnoiu

et al. 2010

J. Am. Soc. Mass Spectrom.

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“…0.25-100 μL/min) were particularly designed in the form of opentubular reactors by the immobilisation of enzymes (i.e. trypsin, pepsin) in fused silica capillaries [38][39][40]. The microcolumn IMER in present study is also one of the first microIMERs designed in the form of a monolithic structure in a fused silica capillary.…”

Section: Resultsmentioning

confidence: 97%

Synthesis of a monolithic, micro-immobilised enzyme reactor via click-chemistry

2012

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An immobilised enzyme reactor (IMER) in the form of capillary monolith was developed for a micro-liquid chromatography system. The plain monolith was obtained by in situ thermal copolymerisation of glycidyl methacrylate and ethylene dimethacrylate in a fused silica capillary (200 × 0.53 mm ID) by using n-propanol/1,4-butanediol as porogen. The enzyme, α-chymotrypsin (CT), was covalently attached onto the monolith via triazole ring formation by click-chemistry. For this purpose, the monolithic support was treated with sodium azide and reacted with the alkyne carrying enzyme derivative. CT was covalently linked to the monolith by triazole-ring formation. The activity behaviour of monolithic IMER was investigated in a micro-liquid chromatography system by using benzoyl-L-tyrosine ethyl ester (BTEE) as synthetic substrate. The effects of mobile-phase flow rate and substrate feed concentration on the final BTEE conversion were investigated under steady-state conditions. In the case of monolithic IMER, the final substrate conversion increased with increasing feed flow rate and increasing substrate feed concentration. Unusual behaviour was explained by the presence of convective diffusion in the macropores of monolith. The results indicated that the monolithic-capillary IMER proposed for micro-liquid chromatography had significant advantages with respect to particle-based conventional high-performance liquid chromatography-IMERs.

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“…Notwithstanding its clear advantages, SPR suffers from the inherent drawback that no structural information is generated. In this respect, SPR has been combined off-line and on-line with MS [17], off-line or in parallel with HPLC-MS [18], and on-line with HPLC-MS [19,20]. Nevertheless, the above mentioned hyphenated technologies were mostly focused on the macrobiomolecules analysis and the interfaces for on-line technology were complicated.…”

Section: Introductionmentioning

confidence: 96%

Simultaneous ligand fishing and identification of human serum albumin binders from Eucommia ulmoides bark using surface plasmon resonance-high performance liquid chromatography–tandem mass spectrometry

Peng

Zhang

Shi

et al. 2013

Journal of Chromatography B

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Development of an on-line SPR-digestion-nanoLC-MS/MS system for the quantification and identification of interferon-γ in plasma

Cited by 22 publications

References 27 publications

On-line bioaffinity-electrospray mass spectrometry for simultaneous detection, identification, and quantification of protein-ligand interactions

On-line bioaffinity-electrospray mass spectrometry for simultaneous detection, identification, and quantification of protein-ligand interactions

Synthesis of a monolithic, micro-immobilised enzyme reactor via click-chemistry

Simultaneous ligand fishing and identification of human serum albumin binders from Eucommia ulmoides bark using surface plasmon resonance-high performance liquid chromatography–tandem mass spectrometry

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