Pyroglutamate Assay

Pyroglutamate (pE) is a glutamic acid dehydration product formed by dehydration of an amino group of an amino-terminal glutamic acid with a 5-carboxy group in some polypeptides. Scientists have demonstrated that the dehydration reactions causing some potential safety hazards can present both in vitro and in vivo. Meanwhile, these reactions can also affect the efficacy of antibodies. Thus, it’s necessary to evaluate the exposure of pE. Creative Biolabs provides a panel of assays to reduce the negative impact on potential drugs by detecting and analyzing the content of pE.

Introduction

pE is a cyclized derivative formed by dehydration of a side chain carboxyl group of glutamic acid at the N-terminus of some active antibodies with an N-terminal amino group. This non-enzymatic reaction is common and has similar chemical transformations at both the heavy and light chains of the antibody. Scientists believe that the formation of pE is a mechanism for the stabilization of antibodies and peptides in vivo. Such cyclized groups are generally resistant to aminopeptidases and thus can buffer physiological cycling. Currently, Creative Biolabs has developed the following two methods for the detection and analysis of pE:

Fig.1 Pyroglutamate formation and its presence and function in fungal hydrolytic enzymes. (Wu, 2017)

Peptide Mapping
Peptide mapping is the main means of analyzing and identifying protein products and preparations in our company. The results have suggested that it is a highly specific identification method for pE analysis.
- Demonstrate a protein's primary structure (amino acid sequence) includes its N and C terminus.
- Demonstrate the site and proportion of protein modifying groups, such as glycosylation, acetylation, sulfation, and phosphorylation.
- Provide qualitative and quantitative information about the protein degradation products.
- Provide information on protein oxidation and deamination.
Cation Exchange
pE is converted from glutamate, accompanied by a charge change, so it can be predicted and analyzed using the cation exchange method.
- Purification of antibodies containing N-terminal pE.
- Evaluating the ability to convert from glutamic acid to pE.
- Detection of pE content.
Our Highlights
- Multiple testing and evaluation equipment.
- Rich research experience.
- One-stop customization, technical service.
- Accurate data analysis.

Creative Biolabs owns advanced equipment and rich research experience in the forecasting and analysis of pyroglutamate in the drug discovery process. If you want to know more information, please contact us and we will happy to help you.

Reference

Wu, V.W.; et al. Identification of Glutaminyl Cyclase Genes Involved in Pyroglutamate Modification of Fungal Lignocellulolytic Enzymes. American Society for Microbiology. 2017, 8(1): e02231-16.

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