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Imaged capillary isoelectric focusing (icIEF) has made remarkable strides in recent years, presenting the biopharmaceutical industry with a range of innovative solutions to characterize protein charge heterogeneity and streamline the development and manufacturing of biologics. This review highlights the latest advancements in icIEF technology for comprehensive biopharmaceutical characterization. Key innovations, including the development of critical reagents and capillary coatings, advanced icIEF fractionation, cutting-edge icIEF-MS online coupling, and novel applications for protein bio-interactions, are redefining icIEF analytical methodologies and broadening their applicability. These breakthroughs significantly enhance the characterization of complex therapeutic proteins , aiding researchers in mitigating challenges and setbacks during drug development and manufacturing. In describing these advances, we delve into multidisciplinary concepts spanning biology, chemistry , instrument design, and workflow optimization, aiming to inspire further innovations and insights in this dynamic, rapidly evolving field. Furthermore, this review traces the origins and 30-year evolution of icIEF technology by illustrating its continuous progression and expanding impact over time.
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Tao et al. (2025) studied this question.
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