Minimal Proteomic sample preparation (mPOP)

 

mPOP Article Nature Protocol JoVE Protocol

Minimal ProteOmic sample Preparation (mPOP) method for single-cell proteomics

 

Introduction Advantages of mPOP RAW Data Videos

Introduction

A major limitation to applying quantitative LC-MS/MS proteomics to small samples, such as single cells, are the losses incurred during sample cleanup. To relieve this limitation, Minimal ProteOmic sample Preparation (mPOP) uses only mass-spectrometry compatible chemicals and thus obviates cleanup while expediting sample preparation and simplifying its automation. Bulk SILAC samples processed by mPOP or by conventional urea-based methods demonstrated that mPOP results in complete, unbiased cell lysis and accurate relative quantification. mPOP can be easily intergraded with all methods for single-cell proteomics by mass-spectrometry. Details of its integration with SCoPE2 are described in this Nature protocol.

Advantages of mPOP

  1. mPOP uses equipment found in most laboratories
    • Easy to adopt and implement without specialized equipment
  2. mPOP uses only mass-spec compatible chemicals
    • No cleanup steps are needed
  3. mPOP is easy to automate
    • Thus mPOP can prepare hundreds of single cells with little human handling
  4. mPOP uses inexpensive and accessible consumables
    • This keeps costs below 1 USD / single cell.

The least reliable step of mPOP is single cell isolation via FACS sorting. The reliability of this step is dependent on the sorters and operators and can be high with experienced operators. Single cells can also be isolated by laser capture microdissection, manual cell picking, CellenONE, magnetic-activated cell sorting, and other methods. We have observed highly consistent and reliable results from mPOP when isolating single cells with CellenONE.

 

Applications & data

 

 

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About the project

This project on developing an automated sample preparation method for single-cell proteomics was supported by funding from the NIH Director’s Award.