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Measuring Antibody Molecular Weight by SEC-MALS

Molecular weight measurements of an antibody using Viscotek SEC-MALS 20

Abstract

In this application note, a purified polyclonal antibody (IgG) is separated using Size Exclusion Chromatography (SEC) and characterized using the Viscotek SEC-MALS 20, allowing calculation of molecular weight and radius of gyration (Rg).

Introduction

Therapeutic recombinant antibodies, primarily of the Immunoglobulin G isotype (IgG) (figure 1), represent a growing proportion of biopharmaceuticals. There are 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.

Figure 1: Molecular structure of an Immunoglobulin.

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. Sizeexclusion chromatography (SEC) is an established and powerful tool that is commonly implemented to assess the aggregation content of protein formulations.

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