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Related Concept Videos

Sample Preparation for Analysis: Overview01:21

Sample Preparation for Analysis: Overview

Sample preparation is an essential step in the analytical process. It involves preparing a sample so that it can be analyzed accurately. The goal is to extract the analyte, the substance you want to measure, from the sample while removing any components that may interfere with the analysis. Sample preparation techniques vary depending on the physical state of the sample.
Bulk or large solid samples are typically reduced in size using grinding, crushing, or milling techniques to increase the...

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Digital Microfluidics for Automated Proteomic Processing
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Simple and Efficient Microsolid-Phase Extraction Tip-Based Sample Preparation Workflow to Enable Sensitive Proteomic

James C Kostas1, Michal Greguš1, Jan Schejbal1

  • 1Barnett Institute of Chemical and Biological Analysis, Department of Chemistry and Chemical Biology, Northeastern University, 360 Huntington Avenue, Boston, Massachusetts 02115, United States.

Journal of Proteome Research
|February 24, 2021
PubMed
Summary

We developed a novel on-microsolid-phase extraction tip (OmSET) method for efficient proteomic profiling of limited biological samples. This technique enables robust analysis of rare cells and small tissue sections, improving sample preparation efficiency and reducing sample loss.

Keywords:
OmSET (on-microsolid-phase extraction tip)bottom-up nanoLC−MS-based proteomicslabel-free quantitationlimited samplesmicro-SPE tipon-membrane digestionsample preparation

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Area of Science:

  • Biochemistry
  • Proteomics
  • Analytical Chemistry

Background:

  • Proteomic profiling of limited biological samples (e.g., rare cells, tissue sections) is challenging due to inefficient sample preparation and sample loss.
  • Existing methods struggle with low cell numbers, hindering in-depth analysis.
  • Need for improved sample preparation techniques for trace biological materials.

Purpose of the Study:

  • To develop and validate a novel sample preparation method for in-depth proteomic analysis of limited biological samples.
  • To address the limitations of current methods in handling small sample volumes and cell numbers.
  • To enable efficient and reproducible proteomic profiling from minimal starting material.

Main Methods:

  • Development of on-microsolid-phase extraction tip (OmSET) for sample preparation.
  • Minimal sample processing volumes (1-3 μL) within a single microreactor.
  • Assessment of OmSET for nanogram-level tryptic peptides and quantitative analysis of low numbers of human monocytes using label-free quantitation.

Main Results:

  • OmSET is simple, efficient, reproducible, and scalable for processing approximately 200 to 10,000 cells.
  • Achieved excellent correlations between protein abundances and starting material (R² = 0.93) and between replicates (R² = 0.95).
  • Identified approximately 300, 1800, and 2000 protein groups from 10, 100, and 500 cell equivalents, respectively.

Conclusions:

  • OmSET significantly improves proteomic profiling of limited biological samples.
  • The method enables reliable quantitative proteomics from very small cell numbers.
  • OmSET is a valuable tool for analyzing trace biological materials in various research applications.