Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)

The overall goal of this enhanced multiplex method is to increase the number of protein samples that can be analyzed simultaneously while decreasing sample error and instrument time. This method can help answer key questions in the fields of aging, neurodegenerative disease, other age-related diseases, and cancer that are related to pathogenesis, progression, diagnosis, prognosis, and therapeutic targets. The main advantage of this technique is that a comprehensive snapshot of protein changes across many conditions can be generated.

Though this method can provide insight into Alzheimer disease in mouse model, it can also be applied to tissue samples from patient with Alzheimer or a range of other disorders. Generally, individuals new to this method will find it challenging because there are several samples to handle simultaneously in a short time frame. We first had the idea for this method when we recognized that acetylation could be combined with isoberic tagging for protein nitration which motivated us to make the technique work globally for all peptides.