Assessing and monitoring the quality of therapeutic mAb products

mAb analysis by peptide mapping
Alphalyse
Nov 25. 2019

Question:

Which analyses should be part of mAb characterization?

I need to ensure the quality and the consistency of my mAbs. Do you have any advice on which analyses I should conduct for proper mAb characterization?

Answer:

For several years, monoclonal antibodies (mAbs) have been the fastest-growing type of pharmaceutical molecules [1]:

More than half of all new drugs approved from 2015 to 2018 were mAbs [2].

Also, the number of approved mAb biosimilars has increased substantially during the last few years.

Before approval for clinical administration, you must show that you have characterized the biosimilar in detail, just like its originator product [2].

What I suggest

Characterization of mAbs can be pretty complex, though. It is not enough to check for the correct amino acid sequence – you must also analyze the structure of the N-glycans and the proper formation of disulfide bridges [2].

We suggest a comprehensive and robust mAb characterization program based on intact mass analysis and peptide mapping to get the complete picture.

A mAb characterization program often consists of the following steps:

Intact mass analysis

  • Can be used before and after reduction
  • Applicable before and after removal of N-glycans
  • As subunits after IdeS treatment

Peptide mapping

  • Application of single or multiple digests using proteases, such as Trypsin, Chymotrypsin, Asp-N, Glu-C, Lys-C, etc.
  • Digestion under reduced and non-reduced conditions

Data analysis

  • Detailed characterization is based on the experimental data obtained by intact mass analysis and peptide mapping.
  • A high level of protein chemistry expertise is necessary for this process.

This workflow provides a reproducible, accurate, sensitive, and fast structural assessment.

The solution is particularly beneficial to the biopharmaceutical industry. Pharma companies thus use it to control mAb product quality and consistency, batch-to-batch comparability, and process modification.

For more information about characterization analysis, I suggest you visit our page dedicated to antibody analysis. Here you can learn about the different analyses, e.g., assessing modifications like glycosylations and quantifying protein.

Amino Acid analysis

Related blog posts:

References:

[1]          Liu, J.K.H.: “The history of monoclonal antibody development – Progress, remaining challenges, and future innovations,” Annals of Medicine and Surgery, 2014

[2]          Walsh, G.: “Biopharmaceutical benchmarks 2018”, Nature Biotechnology, 2018

More posts about Analysis of Biopharmaceuticals
  • Host Cell Protein monitoring in process development – using mass spec

  • Identification of small host cell proteins (small Mw HCPs)

  • Determine the Molar Extinction Coefficient of protein in 3 small steps

  • Why you need accurate concentration determination of protein standards

  • What is SWATH mass spectrometry and its advantages?

About Alphalyse

Alphalyse uses expertise in the field of protein chemistry and bioinformatics, combined with top-of-the-line mass spectrometry equipment, to provide protein analysis services globally.

Alphalyse aims to deliver high quality data and customer service to all of our clients in a fast and convenient way.

www.alphalyse.com

Newsletter Subscribe to The Alphalyse Newsletter and get more
information on protein analysis methods and applications
© 2022 Alphalyse, Inc.