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EMS0011

Sigma-Aldrich

Low-Artifact Digestion Buffer

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

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Synonym(s):
Hi-Fidelity Peptide Mapping Buffer

form

liquid

suitability

suitable for (peptide mapping)

shipped in

wet ice

storage temp.

2-8°C

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S2454T7777MSKT0002
Sigma-Aldrich

EMS0011

Low-Artifact Digestion Buffer

Sigma-Aldrich

MSKT0002

SOLu-Trypsin Rapid Digestion Kit

storage temp.

2-8°C

storage temp.

room temp

storage temp.

2-8°C

storage temp.

2-8°C

suitability

suitable for (peptide mapping)

suitability

-

suitability

-

suitability

suitable for mass spectrometry

shipped in

wet ice

shipped in

-

shipped in

-

shipped in

wet ice

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 Code

12 - Non Combustible Liquids

WGK

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.

This application note compares three different column chemistries from the BIOshell™ line of columns with superficially porous particles to evaluate their performance in peptide mapping.

Workflows for monoclonal antibody adalimumab characterization ensure drug safety and efficacy through critical quality attribute analysis.

Method development for protein fingerprinting of AAV serotype 5 using both intact mass analysis and peptide mapping, to determine critical quality attributes for gene therapy, utilizing three different columns.

Protocols

An optimized LC-MS/MS based workflow for low artifact tryptic digestion and peptide mapping of monoclonal antibody, adalimumab (Humira) using filter assisted sample preparation (FASP).

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

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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