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EMS0011

Sigma-Aldrich

Low-Artifact Digestion Buffer

Minimizes artifactual deamidation and oxidation during trypsin digestion prior to peptide mapping

Synonym(s):

Hi-Fidelity Peptide Mapping Buffer

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About This Item

UNSPSC Code:
41105331

form

liquid

suitability

suitable for (peptide mapping)

shipped in

wet ice

storage temp.

2-8°C

Related Categories

General description

Low-Artifact Digestion Buffer (EMS0011) is optimized for digestion of therapeutic proteins and antibodies prior to peptide mapping by LC-UV-MS. The buffer has been designed to minimize artifactual methionine oxidation and asparagine deamidation during sample preparation while maintaining effective reduction, alkylation and proteolytic digestion.

Endogenous deamidation and oxidation of protein amino acids can be implicated in and be indicative of many diseases, protein turnover, development, and aging. The introduction of these modifications can often affect biological activity, half-life, and immunogenicity.

Current protein digestion workflows often introduce a significant amount of artifactual deamidation and oxidation which prohibits the measurement of the endogenous levels.

The Low Artifact Digestion Buffer allows protein digestion in less than 6 hours with minimized artifactual modifications such as deamidation and oxidation. The amount of oxidation and deamidation introduced during further downstream processing of biotherapeutic antibodies and proteins can be accurately determined. The most commonly deamidated amino acid is Asn especially when followed by small amino acids such as Gly. The most commonly oxidized amino acids are Met, Cys, and to some extent Trp and His.

Storage Class

12 - Non Combustible Liquids

wgk_germany

nwg

flash_point_f

Not applicable

flash_point_c

Not applicable


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Articles

An optimized peptide mapping protocol using NISTmAb as a model monoclonal antibody, shorter incubation times, and improved digestion buffer to demonstrate minimal artificial asparagine deamidation and methionine oxidation.

Step-by-step workflows for the intact mass analysis, peptide mapping, and N-glycan analysis of the monoclonal antibody― adalimumab, for an accurate characterization of the critical quality attributes (CQAs) to ensure drug safety and efficacy. Read more.

Protocols

A step-by-step protocol for released N-linked glycan analysis of the monoclonal antibody adalimumab, based on UHPLC-FLR-MS and procainamide labeling.

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