A cross-sequence labeling equipment for high-throughput proteome based on pseudo-isobaric dimethyl labeling

Facing the needs of multidimensional and multi-sample proteome quantification in the field of biology and precision medicine , high-throughput quantitative labeling and analysis have become the trend in recent years. A novel cross-sequence labeling equipment for high-throughput proteome based on pseudo-isobaric dimethyl labeling (pIDL-StageTip) was established. The equipment needs only simple devices and centrifugal force. The throughput of quantitative labeling is effectively increased , and the controllability of the two-step labeling reaction time at the terminals of the peptides and the ease of operation are ensured. By optimizing the labeling conditions of NaBD3CN and NaBH3CN under acidic conditions respectively, labeling efficiency of the standard proteolysis achieved 100% and labeling selectivity was more than 95%. Even in the complex human proteome , the labeling efficiency was greater than 99% and the labeling selectivity was 100%. The quantitative method based on the equipment showed high quantitative accuracy and precision. The equipment provides a reliable solution for quantitative proteome labeling with high operability, accuracy, and throughput.