A Novel icIEF Fractionation & Surface Plasmon Resonance Workflow
Countless studies on the charge heterogeneity of biotherapeutics report some variation of the statement “charge heterogeneity can have potential impacts on drug stability, efficacy, and potency.” Charge heterogeneity has therefore become a Critical Quality Attribute (CQA) of biotherapeutic molecules, and a combination of assays is required to ascertain the impact of such variants on the product quality.
In this study, we present a novel approach for determining the impact of charge heterogeneity on the binding kinetics and affinity of biotherapeutics. Using the bispecific antibody Mosunetuzumab and a biosimilar as models, the study combines imaged capillary isoelectric focusing (icIEF) on the MauriceFlex™ system with low-volume, high-throughput binding studies on the Alto™ Digital SPR System (Nicoya Lifesciences). This workflow enables efficient isolation and detailed characterization of individual charge variants, offering a novel solution to traditional challenges in charge variant analysis.
icIEF Fractionation, SPR Workflow Highlights
- Charge Heterogeneity Analysis: icIEF on the MauriceFlex System facilitates high-resolution charge variant analysis and collection of high-purity charge species.
- Low-Volume SPR Binding Assays: With only 2 µL required per sample, the Alto Digital SPR System provides real-time kinetic data, significantly reducing material waste and hands-on time.
- Critical Insights: The SPR data correlated well with LC-MS studies done on the same charge variants and highlighted the critical impact of structural changes of the acidic variant of the biosimilar.
Workflow Benefits
It is a fast, accurate, and scalable method to connect charge heterogeneity with binding potency. By identifying charge species that impact binding affinity, this method can enhance process development and manufacturing control, thus supporting higher-quality therapeutic production.
For biotherapeutic developers seeking reliable methods to characterize charge variants and their impact on functionality, this workflow demonstrates the MauriceFlex and Alto systems as a robust solution. Dive deeper into this study’s findings and learn how these insights can support the development of next-generation biologics.
Webinar: Insights into biomolecular charge variants with icIEF and SPR
Hear from Dr. Chris Heger from Bio-Techne and Dr. Mike Piazza from Nicoya Lifescience demonstrate the simple workflow to provide in-depth characterization of the charge variants of a therapeutic bispecific antibody (BsAb) and a research grade biosimilar using the MauriceFlex system and the Alto Digital SPR instrument. They demonstrate that icIEF-based charge separation and fractionation followed by surface plasmon resonance (SPR) analysis revealed notable differences in binding affinity between the innovator drug and a research-grade biosimilar.
Application note: A Novel icIEF Fractionation & SPR-Based Workflow
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