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The Analytical Scientist / App Notes / 2014 / Controlled Reduction of Disulfide Bonds in Proteins/Peptides using on-line EC/MS

Controlled Reduction of Disulfide Bonds in Proteins/Peptides using on-line EC/MS

11/06/2014

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Summary
  • Fast reduction of S-S bonds in an electrochemical flow-through cell
  • Reagent free, no reducing chemicals, e.g., DTT, TCEP, etc.
  • Ideal for TCEP resistant proteins
  • Automated reduction by on-line EC/MS or LC/EC/MS
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antec app note12414
Introduction

Reduction of disulfide bonds in peptides and proteins prior to MS analysis is done for several reasons. It is one of the steps in the determination of the disulfide bonds positions which is important for understanding the folding processes of a protein. Also disulfide bonds can hinder MS identification and their reduction is necessary for efficient sequence analysis in top-down proteomics, or prior to HDX exchange procedures. We developed an electrochemical (EC) method for the efficient and fast reduction of disulfide bonds in peptides and proteins. The method utilizes an electrochemical flow-through cell. The cell can be directly connected to the ESI-MS for flow injection analysis or after the HPLC column in case of complex sample analysis by LC/EC/MS. A complete or near to complete reduction of the disulfide bonds of the tested proteins and peptides has been demonstrated.

>> Download the full Application Note as PDF

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