Application of Simple Western to analysis of antibody target affinity and specificity

Accurate characterization of monoclonal antibody (mAb) target affinity and specificity is critical for antibody development, validation, and diagnostic applications. The Simple Western™ platforms perform capillary–based immunoassays, are fully automated and provide highly reproducible and reliable quantitation. In this webinar, we demonstrate the use of Simple Western platforms to quantify expression levels of mAb in various cell lines, validate antibody specificity using peptide inhibition assays with results comparable to immunohistochemistry data, estimate and compare mAb clone binding affinities, assess effects of substitution of amino acid residues in mAb epitopes and identify and quantify off-target effects. Simple Western technology offers robust, protein analysis that can accelerate selection and optimization of antibody candidates for further therapeutic development and perform highly precise quantitation of biomarkers.

What you will learn:

• An overview of Simple Western technology and how it enables quantitative peptide inhibition and dose-response workflows/assays to estimate mAb target affinity
• Approaches for defining mAb epitopes using peptide truncation and alanine scanning strategies
• Methods for identifying and quantitatively assessing off‑target antibody binding using Simple Western–based workflows