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Concentration‐dependent coulombic effects in travelling wave ion mobility spectrometry collision cross section calibration

Concentration‐dependent coulombic effects in travelling wave ion mobility spectrometry collision cross section calibration
Concentration‐dependent coulombic effects in travelling wave ion mobility spectrometry collision cross section calibration
Rationale
Travelling wave ion mobility spectrometry (TWIMS) is increasingly being used as a method for calculating the collision cross sections (CCSs) of protein ions. To calculate the CCS values of unknown ions, however, the TWIMS device needs to be calibrated using calibrant proteins of known CCS values. The effect of calibrant protein concentration on the accuracy of the resulting calibration curve has not been explicitly studied so far. We hypothesised that at high protein concentrations the ion density within the TWIMS device will be such that ions will experience space charge effects resulting in deviations, as well as broadening, of ion arrival time distributions (ATDs). Calibration curves using these altered ATDs would therefore result in incorrect CCS values being calculated for the protein ions of interest.

Methods
Three protein CCS calibrants, avidin, bovine serum albumin and β‐lactgobulin, were prepared at different concentrations and used to calculate the CCS of a non‐calibrant protein. Data were collected on a Synapt G1 ion mobility mass spectrometer with a nano‐electrospray ionisation (nESI) source using capillaries prepared in house.

Results
Increasing the concentration of CCS calibrants caused ATD broadening and shifted the ATD peak tops, leading to a significant increase in calculated CCS values.

Conclusions
The concentration of protein calibrants can directly affect the quality of the CCS calibration in TWIMS experiments.
0951-4198
Eldrid, Charles
caf78c85-1eae-4700-814a-01b5f0635a0b
O'Connor, Eloise
7cd2afea-c7cb-4081-a5a3-70ffd451e872
Thalassinos, Konstantinos
75b3f786-6a27-420a-8727-97eafb34c022
Eldrid, Charles
caf78c85-1eae-4700-814a-01b5f0635a0b
O'Connor, Eloise
7cd2afea-c7cb-4081-a5a3-70ffd451e872
Thalassinos, Konstantinos
75b3f786-6a27-420a-8727-97eafb34c022

Eldrid, Charles, O'Connor, Eloise and Thalassinos, Konstantinos (2020) Concentration‐dependent coulombic effects in travelling wave ion mobility spectrometry collision cross section calibration. Rapid Communications in Mass Spectrometry. (doi:10.1002/rcm.8613).

Record type: Article

Abstract

Rationale
Travelling wave ion mobility spectrometry (TWIMS) is increasingly being used as a method for calculating the collision cross sections (CCSs) of protein ions. To calculate the CCS values of unknown ions, however, the TWIMS device needs to be calibrated using calibrant proteins of known CCS values. The effect of calibrant protein concentration on the accuracy of the resulting calibration curve has not been explicitly studied so far. We hypothesised that at high protein concentrations the ion density within the TWIMS device will be such that ions will experience space charge effects resulting in deviations, as well as broadening, of ion arrival time distributions (ATDs). Calibration curves using these altered ATDs would therefore result in incorrect CCS values being calculated for the protein ions of interest.

Methods
Three protein CCS calibrants, avidin, bovine serum albumin and β‐lactgobulin, were prepared at different concentrations and used to calculate the CCS of a non‐calibrant protein. Data were collected on a Synapt G1 ion mobility mass spectrometer with a nano‐electrospray ionisation (nESI) source using capillaries prepared in house.

Results
Increasing the concentration of CCS calibrants caused ATD broadening and shifted the ATD peak tops, leading to a significant increase in calculated CCS values.

Conclusions
The concentration of protein calibrants can directly affect the quality of the CCS calibration in TWIMS experiments.

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More information

Accepted/In Press date: 28 October 2019
e-pub ahead of print date: 9 September 2020

Identifiers

Local EPrints ID: 446803
URI: http://eprints.soton.ac.uk/id/eprint/446803
ISSN: 0951-4198
PURE UUID: d5dc4c21-26aa-4517-9c57-28ec148553e8

Catalogue record

Date deposited: 23 Feb 2021 17:31
Last modified: 14 Sep 2021 20:43

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Contributors

Author: Charles Eldrid
Author: Eloise O'Connor
Author: Konstantinos Thalassinos

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