Two‐Dimensional Tandem Mass Spectrometry for Biopolymer Structural Analysis

Author:

Le MyPhuong T.1ORCID,Holden Dylan T.1ORCID,Manheim Jeremy M.2ORCID,Dziekonski Eric T.1ORCID,Iyer Kiran2ORCID,Graham Cooks R.1ORCID

Affiliation:

1. Purdue University Department of Chemistry West Lafayette IN 47907 USA

2. Merck & Co., Inc. Analytical Research and Development Rahway NJ 07065 USA

Abstract

AbstractBiopolymer analysis, including proteomics and glycomics, relies heavily on the use of mass spectrometry for structural elucidation, including sequence determination. Novel methods to improve sample workup, instrument performance, and data analysis continue to be developed to address shortcomings associated with sample preparation, analysis time, data quality, and data interpretation. Here, we present a new method that couples in‐source collision‐induced dissociation (IS‐CID) with two‐dimensional tandem mass spectrometry (2D MS/MS) as a way to simplify proteomics and glycomics workflows while also providing additional insight into analyte structures over traditional MS/MS experiments. Specifically, IS‐CID is employed as a gas‐phase digestion method, i.e., to break down intact full‐length polysaccharide or peptide ions prior to mass analysis. The resulting mixtures of oligomeric ions are analyzed by 2D‐MS/MS, a technique that allows association of product ions with their precursor ions without isolation of the latter. A novel data analysis strategy is introduced to leverage the second dimension of 2D MS/MS spectra, in which stairstep patterns, representing outputs of a molecule's MSn scans, are extracted for structural interconnectivity information on the oligomer. The results demonstrate the potential applicability of 2D MS/MS strategies to the modern omics workflow and structural analysis of various classes of biopolymers.

Funder

Merck Sharp and Dohme

National Science Foundation

Publisher

Wiley

Subject

General Medicine

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