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Electrospray ionization time of flight esi tof mass spectrometer

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The Electrospray Ionization Time of Flight (ESI-TOF) mass spectrometer is a laboratory instrument that uses electrospray ionization and time-of-flight mass analysis to determine the mass-to-charge ratio of ions in a sample. It is designed to provide accurate and sensitive detection and identification of molecular compounds.

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Lab products found in correlation

Bioconfirm software, by Agilent Technologies (5 mentions) Pngase f, by New England Biolabs (5 mentions) Agilent bioconfirm software, by Agilent Technologies (3 mentions) Protein g hp spintrap, by GE Healthcare (1 mentions) Spintrap, by Cytiva (1 mentions)

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Spectrometers (10 citations)

8 protocols using electrospray ionization time of flight esi tof mass spectrometer

1

Deglycosylation and Mass Spectrometry Analysis

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For antibody or antibody conjugate, 20 μg were deglycosylated with PNGase F (New England Biolabs) under reducing conditions (50 mM DTT in PBS) at 37°C overnight. The samples were analyzed on an Agilent Electrospray Ionization Time of Flight (ESI-TOF) mass spectrometer. Deconvoluted masses were obtained using Agilent BioConfirm Software. The conjugation extent by mass spectrometry was calculated from peak integrations of the corresponding deconvoluted mass peaks.
Nilchan N., Alburger J.M., Roush W.R, & Rader C. (2021). An Engineered Arginine Residue of Unusual pH-Sensitive Reactivity Facilitates Site-Selective Antibody Conjugation. Biochemistry, 60(14), 1080-1087.
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2

Mass Spectrometry Analysis of TVD-Fab

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First, 1 mg/ml of TVD–Fab before and after conjugation was prepared for mass spectrometry and data were obtained on an Agilent Electrospray Ionization Time of Flight (ESI-TOF) mass spectrometer. Deconvoluted masses were obtained using Agilent BioConfirm Software.
Hwang D, & Rader C. (2020). Site-Specific Antibody–Drug Conjugates in Triple Variable Domain Fab Format. Biomolecules, 10(5), 764.
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3

Conjugation and Analysis of DVD-Fab Compounds

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Following conjugation of 5 equivalents of compound 1, compound 2, compound 3, or compound 4 to h38C2_K99C DVD-Fab, free compounds were removed with PD MiniTrap G-25 columns and the conjugated DVD-Fabs were concentrated to 1 mg/mL in PBS by using Amicon Ultra 0.5-mL Centrifugal Filters. Following dilution with water, data were acquired on an Agilent Electrospray Ionization Time of Flight (ESI-TOF) mass spectrometer and deconvoluted masses were analyzed with Agilent BioConfirm Software.
Hwang D., Nilchan N., Park H., Roy R.N., Roush W.R, & Rader C. (2022). Sculpting a uniquely reactive cysteine residue for site-specific antibody conjugation. Bioconjugate chemistry, 33(6), 1192-1200.
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4

Enzymatic Deglycosylation and Mass Spectrometry

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All samples were enzymatically deglycosylated with PNGase F (New England Biolabs) overnight at 37°C under reducing conditions (50-mM DTT, PBS pH 7.4). Data were obtained on an Agilent Electrospray Ionization Time of Flight (ESI-TOF) mass spectrometer. Deconvoluted masses were obtained using Agilent BioConfirm software.
Nilchan N., Li X., Pedzisa L., Nanna A.R., Roush W.R, & Rader C. (2019). Dual-mechanistic antibody-drug conjugate via site-specific selenocysteine/cysteine conjugation. Antibody Therapeutics, 2(4), 71-78.
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5

Enzymatic Deglycosylation and Mass Spectrometry

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All samples were enzymatically deglycosylated with PNGase F (New England Biolabs) overnight at 37ºC under reducing conditions (50 mM DTT, PBS pH 7.4). Data was obtained on an Agilent Electrospray Ionization Time of Flight (ESI-TOF) mass spectrometer. Deconvoluted masses were obtained using Agilent BioConfirm software.
Nilchan N., Li X., Pedzisa L., Nanna A.R., Roush W.R, & Rader C. (2019). Dual-mechanistic antibody-drug conjugate via site-specific selenocysteine/cysteine conjugation. Antibody Therapeutics, 2(4), 71-78.
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6

Deglycosylation and Mass Spectrometry

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All samples were enzymatically deglycosylated with PNGase F (New England Biolabs) overnight at 37 °C under reducing conditions (50 mM DTT in PBS). The enzyme was removed using a Protein G HP SpinTrap (GE Healthcare) according to the manufacturer’s instructions. The samples were reduced again prior to sample analysis (10 mM DTT in PBS). Data were obtained on an Agilent Electrospray Ionization Time of Flight (ESI-TOF) mass spectrometer. Deconvoluted masses were obtained using Agilent BioConfirm Software.
Nanna A.R., Li X., Walseng E., Pedzisa L., Goydel R.S., Hymel D., Burke Jr. T.R., Roush W.R, & Rader C. (2017). Harnessing a catalytic lysine residue for the one-step preparation of homogeneous antibody-drug conjugates. Nature Communications, 8, 1112.
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7

Antibody Deglycosylation and Mass Spectrometry

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Unconjugated and conjugated antibodies at 10 μM were reduced with 50 mM DTT in PBS for 10 min at RT followed by enzymatic deglycosylation with PNGase F (New England Biolabs) overnight at 37°C. Following dilution into water, data were obtained on an Agilent Electrospray Ionization Time of Flight (ESI-TOF) mass spectrometer. Deconvoluted masses were obtained using Agilent BioConfirm Software.
Hwang D., Tsuji K., Park H., Burke TR J.r, & Rader C. (2019). Site-Specific Lysine Arylation as an Alternative Bioconjugation Strategy for Chemically Programmed Antibodies and Antibody-Drug Conjugates. Bioconjugate chemistry, 30(11), 2889-2896.
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8

Conjugation and Mass Spectrometry of DVD-Fab

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Following conjugation of 5 and 10 equivalents of phenylglyoxal-TAMRA (compound 1) or 5 equivalents of phenylglyoxal-MMAF (compound 4) to h38C2_Arg DVD-Fab as described above, illustra NAP-5 Columns were used to remove free compound and the conjugated DVD-Fab was concentrated with Amicon Ultra 0.5-mL Centrifugal Filters to 1 mg/mL in PBS (pH 7.4). Following dilution into water, data were obtained on an Agilent Electrospray Ionization Time of Flight (ESI-TOF) mass spectrometer. Deconvoluted masses were obtained using Agilent BioConfirm Software.
Hwang D., Nilchan N., Nanna A.R., Li X., Cameron M.D., Roush W.R., Park H, & Rader C. (2019). Site-Selective Antibody Functionalization via Orthogonally Reactive Arginine and Lysine Residues. Cell chemical biology, 26(9), 1229-1239.e9.
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