SNaPP: Simplified Nano-Proteomics Platform for reproducible global proteomic analysis of nanogram protein quantities (original) (raw)

Endocrinology, 2016

Abstract

Global proteomic analyses of complex protein samples in nanogram quantities require a fastidious approach to achieve in-depth protein coverage and quantitative reproducibility. Biological samples are often severely mass-limited and can preclude the application of more robust bulk sample processing workflows. In this study, we present a system that minimizes sample handling by using online immobilized trypsin digestion and solid phase extraction to create a simple, sensitive, robust, and reproducible platform for the analysis of nanogram-size proteomic samples. To demonstrate the effectiveness of our Simplified Nano-Proteomics Platform (SNaPP), we utilized the system to analyze preimplantation blastocysts collected on day 4 of pregnancy by flushing the uterine horns with saline. For each of our 3 sample groups, blastocysts were pooled from 3 mice resulting in 22, 22, and 25 blastocysts, respectively. The resulting proteomic data provides novel insight into mouse blastocyst protein ex...

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