Cookies

Like most websites The Analytical Scientist uses cookies. In order to deliver a personalized, responsive service and to improve the site, we remember and store information about how you use it. Learn more.
Techniques & Tools Sample Preparation, Mass Spectrometry, Technology, Data Analysis, Pharma & Biopharma, Clinical, Proteomics, Liquid Chromatography, Gas Chromatography, Thin Layer Chromatography, Preparative/Process Chromatography

Measuring Antibody Molecular Weight by SEC-MALS

sponsored by Malvern Panalytical

Figure 1: Molecular structure of an Immunoglobulin

Introduction

Therapeutic recombinant antibodies, primarily of the Immunoglobulin G isotype (IgG) (figure 1), represent a growing proportion of biopharmaceuticals.  There is an ever increasing number of approved antibody therapies available to treat a variety of modern-day diseases such as cancer, rheumatoid arthritis and diabetes. With many more candidate IgG's in the biopharmaceutical development pipeline, the pressure to accurately understand and develop these candidates becomes more and more important.

Protein aggregation has long been established as a major concern in the biopharmaceutical industry.  Proteins have a limited shelf-life and precipitate over time, with strong evidence that the presence of protein aggregates in the bloodstream can regularly stimulate the active/adaptive immune response. Size-exclusion chromatography (SEC) is an established and powerful tool that is commonly implemented to assess the aggregation content of protein formulations.

SEC separates proteins by size, and is frequently employed to measure molecular weight and identify aggregate sample proportions. While most SEC systems use a single detector (such as an ultraviolet absorbance detector), the addition of a light scattering detector allows direct measurement of protein molecular weights and is used to differentiate between monomer, dimer and aggregate independent of retention volume. Light scattering is substantially more sensitive to aggregate formation than absorbance measurements and allows detection of aggregates that may otherwise be missed. If the light scattering detector employed is a multi angle light scattering (MALS), then it can also be used to measure the radius of gyration (Rg) of the large aggregates that scatter light anisotropically (unevenly with respect to angle), which offers further structural/shape information.

The Viscotek SEC-MALS 20 system (figure 2) is a 20 angle light scattering device, capable of making measurements of protein and protein aggregate molecular weight, independent of elution volume.  

Read the full article now

Log in or register to read this article in full and gain access to The Analytical Scientist’s entire content archive. It’s FREE and always will be!

Login

Or register now - it’s free and always will be!

You will benefit from:

  • Unlimited access to ALL articles
  • News, interviews & opinions from leading industry experts
  • Receive print (and PDF) copies of The Analytical Scientist magazine
Register

Or Login via Social Media

By clicking on any of the above social media links, you are agreeing to our Privacy Notice.

Newsletter

Send me the latest from The Analytical Scientist.

Sign up now

Register to The Analytical Scientist

Register to access our FREE online portfolio, request the magazine in print and manage your preferences.

You will benefit from:

  • Unlimited access to ALL articles
  • News, interviews & opinions from leading industry experts
  • Receive print (and PDF) copies of The Analytical Scientist magazine

Register