Captivesprayer

Manufactured by Bruker

The CaptiveSprayer is a laboratory equipment product. It is a specialized instrument designed for controlled sample introduction in mass spectrometry analysis.

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Lab products found in correlation

Halo fused core particles, by Merck Group (1 mentions) Acclaim pepmap 100 c18 trap column, by Thermo Fisher Scientific (1 mentions) Maxis impact, by Bruker (1 mentions) Maxis hd, by Bruker (1 mentions) Acclaim pepmap 100 trap column, by Thermo Fisher Scientific (1 mentions) Ultimate 3000 rslcnano system, by Thermo Fisher Scientific (1 mentions) Nanobooster, by Bruker (1 mentions)

2 protocols using captivesprayer

Proteomic Analysis of IgG and PR3-ANCA by nanoLC-MS

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The digested samples were analyzed by nanoLC-reversed phase (RP)-electrospray ionization (ESI) – quadrupole time-of-flight (qTOF)-MS on an Ultimate 3000 HPLC system (Dionex/Thermo Scientific, Sunnyvale, CA) coupled to a MaXis Impact (Bruker Daltonics, Bremen, Germany). The samples were concentrated on a Dionex Acclaim PepMap100 C18 trap column (particle size 5 μm, internal diameter 300 μm, length 5 mm) and separated on an Ascentis Express C18 nano column (2.7 μm HALO fused core particles, internal diameter 75 μm, length 50 mm; Supelco, Bellefonte, PA). The following linear gradient was applied, with solvent A consisting of 0.1% trifluoroacetic acid (TFA; Fluka) and solvent B of 95% acetonitrile (Biosolve, Valkenswaard, The Netherlands): t = 0 min, 3% solvent B; t = 2, 6%; t = 4.5, 18%; t = 5, 30%; t = 7, 30%; t = 8, 1%; t = 10.9, 1%. The sample was ionized in positive ion mode with a CaptiveSprayer (Bruker Daltonics) at 1100 V. A nanoBooster (Bruker Daltonics) was used to enrich the nitrogen gas with acetonitrile to enhance the ionization efficacy. A mass spectrum was acquired every second (frequency of 1 Hz), with the ion detection window set at m/z 550–1800. In between every 12 measurements an external IgG standard was run. A mass spectrum of both total IgG and PR3-ANCA can be seen in Fig. 1.

Kemna M.J., Plomp R., van Paassen P., Koeleman C.A., Jansen B.C., Damoiseaux J.G., Cohen Tervaert J.W, & Wuhrer M. (2017). Galactosylation and Sialylation Levels of IgG Predict Relapse in Patients With PR3-ANCA Associated Vasculitis. EBioMedicine, 17, 108-118.

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LC-MS/MS Proteomics Workflow

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Sample (preparation mentioned above) was injected into an Ultimate 3000 RSLCnano system (Thermo Scientific, Breda, the Netherlands) coupled to a quadrupole-TOF-MS (MaXis HD; Bruker Daltonics, Bremen, Germany). The LC system was equipped with an Acclaim PepMap 100 trap column (particle size 5 μm, pore size 100 Å, 100 μm × 20 mm, Thermo Scientific) and an Acclaim PepMap C18 nano analytical column (particle size 2 μm, pore size 100 Å, 75 μm × 150 mm, Thermo Scientific). The samples were loaded and washed on the trap column for 2 min at 15 μl/min with 0.1% FA in water. A mixture of solvent A (0.1% FA in water) and solvent B (95% acetonitrile, 0.1% FA) was applied with a constant flow of 0.7 μL/min using a linear gradient: t(min) = 0, %B = 1; t = 5, %B = 1; t = 30, %B = 50; with washing and equilibration starting at t = 31, %B = 70; t = 35, %B = 70; t = 36, %B = 1; t = 70, %B = 1. The sample was ionized in positive-ion mode using a CaptiveSprayer (Bruker Daltonics) electrospray source at 1250 V. A nanoBooster (Bruker Daltonics) was used to enrich the nitrogen gas with acetonitrile to enhance ionization efficiency (0.2 bar). Mass spectra were acquired with a frequency of 1 Hz and the MS ion detection window was set at mass-to-charge ratio (m/z) 550–1,800.

Holborough-Kerkvliet M.D., Mucignato G., Moons S.J., Psomiadou V., Konada R.S., Pedowitz N.J., Pratt M.R., Kissel T., Koeleman C.A., Tjokrodirijo R.T., van Veelen P.A., Huizinga T., van Schie K.A., Wuhrer M., Kohler J.J., Bonger K.M., Boltje T.J, & Toes R.E. (2023). A photoaffinity glycan-labeling approach to investigate immunoglobulin glycan-binding partners. Glycobiology, 33(9), 732-744.

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