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Formaldehyde derivatization, a Side Reaction During Filter-Aided Sample Preparation

ABSTRACT: The filter-aided sample preparation (FASP) method has been commonly used for proteomic sample preparation due to its high efficiency and convenience. Herein, we report an overlooked side reaction during FASP method, resulting in a mass increment of 12 Da. We confirmed that the side reaction is caused by formaldehyde released from the spin filter and devel-oped a pretreatment step to reduce the side reaction. We further examined spin filters from different vendors and offered suggestions for different experimental purpose. Our findings would benefit the deep screening of labeling proteomics analy-sis and offer a new direction to improve performance of filter devices used for different biological and proteomics studies.

INSTRUMENT(S): Orbitrap Fusion Lumos, LTQ Orbitrap Elite

ORGANISM(S): Caenorhabditis Elegans

TISSUE(S): Whole Body

SUBMITTER: Zhen Wu

LAB HEAD: Xumin Zhang

PROVIDER: PXD018737 | Pride | 2020-08-20

REPOSITORIES: Pride

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Formaldehyde Derivatization: An Unexpected Side Reaction During Filter-Aided Sample Preparation.

Tang Langlang L Wu Zhen Z Wang Jie J Zhang Xumin X

Analytical chemistry 20200828 18

The filter-aided sample preparation (FASP) method has been commonly used for proteomic sample preparation due to its high efficiency in removing impurities. Herein, we report an overlooked +12 Da side modification during FASP method using Microcon spin filters. We confirmed that the side modification is caused by formaldehyde released from the spin filter and found that the side modification leads to 10.5% and 9.5% loss in proteome-level peptide and protein identification, respectively. We evalu ...[more]

PMID: 32786431

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