High-throughput bioassays using "dip-and-go" multiplexed electrospray mass spectrometry.

A multiplexed system based on inductive nanoelectrospray mass spectrometry (nESI-MS) has been developed for high-throughput screening (HTS) bioassays. This system combines inductive nESI and field amplification micro-electrophoresis to achieve a "dip-and-go" sample loading and purification strategy that enables nESI-MS based HTS assays in 96-well microtiter plates. The combination of inductive nESI and micro-electrophoresis makes it possible to perform efficient in situ separations and clean-up of biological samples. The sensitivity of the system is such that quantitative analysis of peptides from 1-10,000 nM can be performed in a biological matrix. A prototype of the automation system has been developed to handle 12 samples (one row of a microtiter plate) at a time. The sample loading and electrophoretic clean-up of biosamples can be done in parallel within 20 s followed by MS analysis at a rate of 1.3 to 3.5 s/sample. The system was used successfully for the quantitative analysis of BACE1-catalyzed peptide hydrolysis, a prototypical HTS assay of relevance to drug discovery. IC 50 values for this system were in agreement with LC-MS but recorded in times more than an order of magnitude shorter.

[1]  Robert Ries,et al.  Establishing MALDI-TOF as Versatile Drug Discovery Readout to Dissect the PTP1B Enzymatic Reaction , 2018, SLAS discovery : advancing life sciences R & D.

[2]  R. Cooks,et al.  Electrophoretic Desalting To Improve Performance in Electrospray Ionization Mass Spectrometry. , 2018, Analytical chemistry.

[3]  Gongyu Li,et al.  In Situ Living Cell Protein Analysis by Single-Step Mass Spectrometry. , 2018, Analytical chemistry.

[4]  J Willem M Nissink,et al.  Seven Year Itch: Pan-Assay Interference Compounds (PAINS) in 2017—Utility and Limitations , 2017, ACS chemical biology.

[5]  Xiang Fang,et al.  Boosting the Signal Intensity of Nanoelectrospray Ionization by Using a Polarity-Reversing High-Voltage Strategy. , 2017, Analytical chemistry.

[6]  Hongying Zhu,et al.  Single-neuron identification of chemical constituents, physiological changes, and metabolism using mass spectrometry , 2017, Proceedings of the National Academy of Sciences.

[7]  J. Pawliszyn,et al.  Open Port Probe Sampling Interface for the Direct Coupling of Biocompatible Solid-Phase Microextraction to Atmospheric Pressure Ionization Mass Spectrometry. , 2017, Analytical chemistry.

[8]  Thomas L. Fillmore,et al.  Capillary Electrophoresis-Nanoelectrospray Ionization-Selected Reaction Monitoring Mass Spectrometry via a True Sheathless Metal-Coated Emitter Interface for Robust and High-Sensitivity Sample Quantification. , 2016, Analytical chemistry.

[9]  Melanie V. Leveridge,et al.  The Evolution of MALDI-TOF Mass Spectrometry toward Ultra-High-Throughput Screening: 1536-Well Format and Beyond , 2016, Journal of biomolecular screening.

[10]  Jonathan Wingfield,et al.  Novel Acoustic Loading of a Mass Spectrometer , 2016, Journal of laboratory automation.

[11]  Shirly Pinto,et al.  Identification of DGAT2 Inhibitors Using Mass Spectrometry , 2016, Journal of biomolecular screening.

[12]  W. Xu,et al.  Pulsed Direct Current Electrospray: Enabling Systematic Analysis of Small Volume Sample by Boosting Sample Economy. , 2015, Analytical chemistry.

[13]  R. Cooks,et al.  On-demand ambient ionization of picoliter samples using charge pulses. , 2015, Angewandte Chemie.

[14]  J. Hermes,et al.  Use of High-Throughput Mass Spectrometry to Reduce False Positives in Protease uHTS Screens , 2015, Journal of biomolecular screening.

[15]  Liang Qiao,et al.  On-chip spyhole mass spectrometry for droplet-based microfluidics. , 2014, Angewandte Chemie.

[16]  Shuo Han,et al.  Rapid removal of matrices from small-volume samples by step-voltage nanoelectrospray. , 2013, Angewandte Chemie.

[17]  H. Girault,et al.  Electrostatic-spray ionization mass spectrometry. , 2012, Analytical chemistry.

[18]  John P. Overington,et al.  ChEMBL: a large-scale bioactivity database for drug discovery , 2011, Nucleic Acids Res..

[19]  R. Cooks,et al.  Induced nanoelectrospray ionization for matrix-tolerant and high-throughput mass spectrometry. , 2011, Angewandte Chemie.

[20]  R. Cooks,et al.  Synchronized inductive desorption electrospray ionization mass spectrometry. , 2011, Angewandte Chemie.

[21]  C. Jonsson,et al.  A cell-based luminescence assay is effective for high-throughput screening of potential influenza antivirals. , 2007, Antiviral research.

[22]  Y. Lam,et al.  Assessment of Joule heating and its effects on electroosmotic flow and electrophoretic transport of solutes in microfluidic channels , 2006, Electrophoresis.

[23]  D. Kassel.  Combinatonial chemistry and mass spectrometry in the 21st century drug discovery laboratory. , 2001, Chemical reviews.

[24]  N F de Rooij,et al.  Sample preconcentration by field amplification stacking for microchip‐based capillary electrophoresis , 2001, Electrophoresis.

[25]  J. Drews.  Drug discovery: a historical perspective. , 2000, Science.

[26]  J. Ramsey,et al.  Subattomole-Sensitivity Microchip Nanoelectrospray Source with Time-of-Flight Mass Spectrometry Detection. , 1999, Analytical chemistry.

[27]  John F. Hartwig,et al.  A Fluorescence-Based Assay for High-Throughput Screening of Coupling Reactions. Application to Heck Chemistry , 1999 .

[28]  M. Karas,et al.  High-sensitivity analysis of neutral underivatized oligosaccharides by nanoelectrospray mass spectrometry. , 1997, Analytical chemistry.

[29]  M. Wilm,et al.  Analytical properties of the nanoelectrospray ion source. , 1996, Analytical chemistry.

[30]  D. Burgi,et al.  Optimization in sample stacking for high-performance capillary electrophoresis , 1991 .

[31]  R. Chien,et al.  Electroosmotic properties and peak broadening in field-amplified capillary electrophoresis , 1991 .

[32]  F. Everaerts,et al.  High-performance zone electrophoresis , 1979 .