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AccuMAP™ Low pH Protein Digestion Kit

Suppress Artificial Deamidation and Disulfide Bond Scrambling in Protein Sample Prep

  • Sample preparation completed in 4.5–5 hours
  • Highly reproducible digestion results
  • Use for reduced and nonreduced proteins

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Catalog number selected: VA1040

$ 750.00 Your price: Log In

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AccuMAP™ Low pH Protein Digestion Kit
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$ 750.00
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Suppress Nonenzymatic PTMs During Protein Sample Prep

Nonenzymatic posttranslational modifications (PTMs) spontaneously occur in biotherapeutic proteins during manufacturing and storage. The major nonenzymatic PTMs are deamidation, disulfide bond scrambling and oxidation. These modifications can affect efficacy and stability of biotherapeutic proteins and are the subject of careful monitoring.

Nonenzymatic PTMs can also be induced during protein sample preparation for peptide mapping and can compromise analysis. Major causes of nonenzymatic PTMs induced during sample preparation include alkaline pH as well as excipients and impurities that have protein oxidizing activity.

Suppress Artificial Nonenzymatic PTMs

The AccuMAP™ Low pH Protein Digestion Kits suppress artificial nonenzymatic PTMs, as sample preparation is performed at low pH under either reducing or nonreducing conditions.

13959MA-W
Schematic diagram of sample preparation by digestion performed with AccuMAP™ Low pH Protein Digestion Kit.

Protein Digestion at Low pH

Trypsin and other proteases commonly used in peptide mapping sample preparation favor alkaline pH in order to efficiently digest proteins. To avoid artificial nonenzymatic PTMs induced at these conditions, we developed trypsin digestion at low pH. To restore trypsin cleavage efficiency at lysine sites we supplemented trypsin with a special, low pH-resistant recombinant Lys-C (rLys-C) protease. By supplementing trypsin with low pH resistant rLys-C we achieved efficient tryptic digestion at low pH.

13961MA-W
Efficient digestion with modified trypsin and AccuMAP™ Low pH Resistant rLys-C at low pH. Rituximab was digested according to a conventional protocol (at pH 8) or with the AccuMAP™ Low pH Protein Digestion Kit at low pH and analyzed by LC/MS. The data show that digestion at low pH was comparable to digestion at conventional conditions.

Suppression of Deamidation and Disulfide Bond Scrambling

13962MA-W-a-2
13962MA-W-b-2

Complete suppression of deamidation and disulfide bond scrambling in IgG digested with AccuMAP™ Low pH Protein Digestion Kit. Figure shows extracted ion chromatogram of a GLEWIGAIYPGNGDTSYNQK peptide from Rituximab antibody digested at conventional conditions (pH 8) and at low pH (with AccuMAP™ Low pH Protein Digestion Kit). The data show suppression of deamidation in the upper panel and the complete suppression of disulfide bond scrambling in the lower panel.

Minimal Baseline Noise in Tryptic Digests

13963MA-W-a-2
13963MA-W-b-2

UV-HPLC chromatograms of Panitumumab digests. Panitumumab was predigested with AccuMAP™ Low pH Resistant rLys-C, then the reaction was diluted and digestion was completed by incubation with trypsin for 3 hours (upper panel) or overnight (lower panel). In this experiment we used a 1:5 trypsin:protein ratio. Note the accumulation of baseline noise for the overnight digest shown in the lower panel.

Study Compares Biosimilar Monoclonal to Original Antibody Remicade

In April 2016, the FDA approved the first biosimilar monoclonal antibody Remsima, developed from Remicade (infliximab, Janssen). Biosimilars provide significant cost savings for patients, but the potential differences between innovative drug and inexpensive duplicate raise questions regarding product usability. Read about the authors’ comparison of Remicade and Remsima using native mass spectrometry, ion mobility and quantitative peptide mapping, including the use of the Promega Low pH Protein Digestion Kit.

A Multidimensional Analytical Comparison of Remicade and the Biosimilar Remsima. Pisupati, et al. (2017)  ACS Anal. Chem. 89, 4838.

Read the Article

Specifications

You are viewing: VA1040 Change Configuration

What's in the box?

Item Part # SizeAvailable Separately

AccuMAP™ Modified Trypsin Solution

V528C 2 × 60μg View Product

AccuMAP™ Denaturing Solution

VA100A 1 × 1ml View Product

AccuMAP™ 10X Low pH Reaction Buffer

VA101A 1 × 1ml View Product

AccuMAP™ 100X Oxidation Suppressant

VA102A 1 × 50μl View Product

AccuMAP™ Low pH Resistant rLys-C Solution

VA103A 2 × 60μg View Product

TCEP

VB100A 3 × 15mg View Product

Iodoacetamide

VB101A 3 × 15mg View Product

NEM

VB102A 2 × 15mg

SDS

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Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

AA

Specifications

You are viewing: VA1050 Change Configuration

What's in the box?

Item Part # SizeAvailable Separately

AccuMAP™ Modified Trypsin Solution

V528D 3 × 400μg View Product

AccuMAP™ Denaturing Solution

VA100B 2 × 5ml View Product

AccuMAP™ 10X Low pH Reaction Buffer

VA101B 2 × 5ml View Product

AccuMAP™ 100X Oxidation Suppressant

VA102B 2 × 250μl View Product

AccuMAP™ Low pH Resistant rLys-C Solution

VA103B 3 × 400μg View Product

TCEP

VB100A 5 × 15mg View Product

Iodoacetamide

VB101A 5 × 15mg View Product

NEM

VB102A 4 × 15mg

SDS

Choose language:

Certificate of Analysis

Search for Specific Certificate:

View more results
View more results
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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

AA
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