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High-throughput analysis of sub-visible mAb aggregate particles using automated fluorescence microscopy imaging
Albert Jesuran Paul1, Fabian Bickel2,3, Martina Röhm2,3
1Institute of Applied Biotechnology, Biberach University of Applied Sciences, Hubertus-Liebrecht-Strasse 35, 88400, Biberach, Germany. paul@hochschule-bc.de.
Analytical and Bioanalytical Chemistry
|April 28, 2017
Summary
Protein aggregation analysis is crucial for drug safety and efficacy. A new automated fluorescence microscopy method detects and quantifies a broad range of protein aggregates, improving therapeutic protein production.
Area of Science:
- Biopharmaceutical analysis
- Protein aggregation science
- Analytical chemistry
Background:
- Protein aggregation is a critical issue in therapeutic protein production, affecting drug efficacy and patient safety.
- Current methods for analyzing protein aggregates have limitations in size range, quantification, and automation.
- There is a need for a high-throughput, broadly applicable method for detecting and classifying higher molecular weight (HMW) species.
Purpose of the Study:
- To develop and validate a novel, automated method for detecting and quantifying a wide range of protein aggregates.
- To address the limitations of existing analytical techniques for higher molecular weight (HMW) species.
- To enable high-throughput analysis and classification of protein aggregates in therapeutic proteins.
Main Methods:
- Utilized automated fluorescence microscope imaging (aFMI) combined with fluorescent dyes for protein aggregate labeling.
- Employed 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonate (Bis-ANS) as a fluorescent dye for monoclonal antibody (mAb) aggregate detection.
- Validated the novel method against established techniques like SE-HPLC, UV-Vis spectroscopy, and dynamic light scattering.
Main Results:
- The developed aFMI method successfully detected protein aggregates in the size range of 1 to 1000 μm.
- The method enabled reliable quantification and classification of various HMW species of monoclonal antibodies (mAbs).
- The approach demonstrated high-throughput compatibility and adaptability for different aggregate types induced by various stresses.
Conclusions:
- The novel aFMI method offers a fast, automated, and high-throughput solution for analyzing protein aggregates across a broad size spectrum.
- This technique overcomes limitations of traditional methods, allowing for comprehensive detection, quantification, and classification of HMW species.
- The fluorescent dye-based aFMI approach provides a versatile platform for improving therapeutic protein quality control and safety.
