Rapid Removal of IgG1 Carryover on Protease Column Using Protease-Safe Wash Solutions Delivered with LC Pump for HDX-MS Systems

Published

December 23, 2024

Author(s)

Kyle Anderson, Sandeep Kumar

Abstract

Sample carryover is a common problem in hydrogen-deuterium exchange mass spectrometry, particularly because immobilized protease columns cannot withstand the high organic solvent concentrations typically used in liquid chromatography mass spectrometry (LC-MS) for cleaning. Conventional cleaning methods using injections of guanidine HCl still suffer from carryover and may require four blanks after each sample run to fully remove carryover. We have implemented an additional LC pump to deliver customized wash solutions to protease and enzyme columns and the associated LC capillaries to eliminate carryover. Pump-based washing using the mild wash solutions tested herein were able to fully remove carryover with only one blank run. FOS-choline-12 was found to be the most effective component in wash solutions and even performed well alone at 0.1 % concentration. Since the protease column washing is performed concurrently during the analytical gradient in sample runs and since blank runs could be reduced from four to one, total run time could be reduced by up to 60 %. Savings in total run time could more than double the productivity of data acquisition, which is imperative for pandemic preparedness and for acceleration of biotherapeutics development.

Citation

Journal of the American Society for Mass Spectrometry

Volume

36

Issue

2

Pub Type

Journals

Download Paper

https://doi.org/10.1021/jasms.4c00411

Local Download

Keywords

HDX-MS, carryover, NISTmAb

Citation

Anderson, K. and Kumar, S. (2024), Rapid Removal of IgG1 Carryover on Protease Column Using Protease-Safe Wash Solutions Delivered with LC Pump for HDX-MS Systems, Journal of the American Society for Mass Spectrometry, [online], https://doi.org/10.1021/jasms.4c00411, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=958601 (Accessed October 14, 2025)