Quick Antibody Off-target Binding Assessment Via High-Throughput SPR

Key Takeaways

• Carterra has enabled off-target testing on up to 384-antibodies simultaneously against large panels of polyreactivity reagents.
• Up to 1152 antibodies can be tested in a single unattended run.
• Polyreactive agents with diverse biophysical properties exhibit no non-specific binding to Carterra sensor chips allowing for the broadest paneling with minimal interference.

Introduction

Antibody off-target assessment, or polyreactivity assessment, is a crucial test to ensure the specificity and safety of therapeutic antibodies. Polyreactivity testing involves evaluating whether an antibody binds specifically to unintended targets. Off-target binding and poor specificity could lead to adverse effects including triggering unwanted immune responses, which can lead to tissue damage or other toxic effects. Off-target binding can also lead to poor pharmacokinetics, where the antibody is sequestered broadly, reducing its availability to bind the intended target [1,2,3,4]. Addressing off-target binding is crucial for the successful application of antibody therapeutics, and by using high-throughput methods, these tests can be incorporated early in the candidate selection process [5, 6, 7].

Here, we describe a high-throughput surface plasmon resonance assay to assess antibody off-target binding, which can be integrated as part of the drug discovery process. The Carterra LSA monitors the SPR signals of up to 384 ligands in a single array, thus allowing the off-target binding profile of a panel of antibodies to be determined against panels of reagents. In this study we chose two common polyreactivity agents, DNA and lipopolysaccharide (LPS). Their particular biophysical and biochemical properties, such as enrichment in negative charge and amphipathic properties, make them both relevant to polyreactivity testing. Additional reagents can be easily added to expand the breadth of the polyreactivity profile.

Posted by Judicaël Parisot, PhD