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

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 evaluated different pretreatment procedures to reduce the side reaction. Furthermore, on the basis of the evaluation results of different brands of spin filters, we recommend Nanosep spin filters for different proteomic studies, especially for amine-labeling proteomic studies. Our results would benefit researchers employing the spin filters to improve their results and also help spin filter manufacturers to improve the product quality. Data are available via ProteomeXchange with identifier PXD018737.