Mass Spectrometric Distinction of In-Source and In-Solution Pyroglutamate and Succinimide in Proteins: A Case Study on rhG-CSF

Formation of cyclic intermediates involving water or ammonia loss is a common occurrence in any reaction involving terminal amines or hydroxyl group containing species. Proteins that have both these functional groups in abundance are no exception, and presence of amino acids such as asparagine, glut...

Full description

Saved in:
Bibliographic Details
Published in:Journal of the American Society for Mass Spectrometry 2013-02, Vol.24 (2), p.202-212
Main Authors: Kumar, Mukesh, Chatterjee, Amarnath, Khedkar, Anand P., Kusumanchi, Mutyalasetty, Adhikary, Laxmi
Format: Article
Language:English
Subjects:
Abundance
Amines
Amino acids
Ammonia
Analytical Chemistry
Bioinformatics
Biotechnology
Chemistry
Chemistry and Materials Science
Functional groups
Granulocyte Colony-Stimulating Factor - chemistry
Humans
Impurities
Mass spectrometry
Mass Spectrometry - methods
Models, Molecular
Organic Chemistry
Peptide Fragments - analysis
Peptide Fragments - classification
Proteins
Proteomics
Pyrrolidonecarboxylic Acid - analysis
Pyrrolidonecarboxylic Acid - chemistry
Recombinant Proteins - chemistry
Research Article
Spectrometry
Succinimides - analysis
Succinimides - chemistry
Trypsin - chemistry
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Formation of cyclic intermediates involving water or ammonia loss is a common occurrence in any reaction involving terminal amines or hydroxyl group containing species. Proteins that have both these functional groups in abundance are no exception, and presence of amino acids such as asparagine, glutamines, aspartic acids, and glutamic acids aid in formation of such intermediates. In the biopharma scenario, such intermediates lead to product- or process-related impurities that might be immunogenic. Mass spectroscopy is a powerful technique that is used to decipher the presence and physicochemical characteristics of such impurities. However, such intermediates can also form in situ during mass spectrometric analysis. We present here the detection of in-source and in-solution formation of succinimide and pyroglutamate in the protein granulocyte colony stimulating factor. We also propose an approach for quick differentiation of such in-situ species from the tangible impurities. We believe that this will not only reduce the time spent in unambiguous identification of succinimide- and/or pyroglutamate-related impurity in bio-pharmaceutics but also provide a platform for similar studies on other impurities that may form due to stabilized intermediates.
ISSN:1044-0305
1879-1123
DOI:10.1007/s13361-012-0531-7