Mid-infrared spectroscopy-based antibody aggregate quantification in cell culture fluids. 2013

Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Germany. florian.capito@external.merckgroup.com

Therapeutic antibody purification involves several steps which potentially induce antibody aggregation. Currently, aggregate monitoring mainly employs chromatographic, SDS-PAGE and light scattering techniques. In this study, the feasibility of mid-infrared spectroscopy (MIR) for the quantification of soluble antibody aggregates was investigated. Several multivariate models were evaluated to quantify antibody aggregation in chromatography elution streams and in clarified CHO cell culture supernatants (a surrogate for bioreactor output). A general model was established that is applicable for aggregate quantification directly from different cell culture solutions. Real-process samples and process-sample mimics were used to verify the general aggregate quantification model using two different antibodies. Results showed good prediction ability down to 1% aggregate content. Together with recently published results using MIR for host cell protein and target protein quantification, the results presented here indicate that MIR could provide multi-parameter process information from a single, fast, cost-effective and straightforward measurement. In conclusion, our study demonstrates that MIR is suitable for aggregate quantification in therapeutic antibody purification processes.

UI MeSH Term Description Entries
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D003412 Cricetulus A genus of the family Muridae consisting of eleven species. C. migratorius, the grey or Armenian hamster, and C. griseus, the Chinese hamster, are the two species used in biomedical research. Hamsters, Armenian,Hamsters, Chinese,Hamsters, Grey,Armenian Hamster,Armenian Hamsters,Chinese Hamster,Chinese Hamsters,Grey Hamster,Grey Hamsters,Hamster, Armenian,Hamster, Chinese,Hamster, Grey
D006224 Cricetinae A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS. Cricetus,Hamsters,Hamster
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D001709 Biotechnology Body of knowledge related to the use of organisms, cells or cell-derived constituents for the purpose of developing products which are technically, scientifically and clinically useful. Alteration of biologic function at the molecular level (i.e., GENETIC ENGINEERING) is a central focus; laboratory methods used include TRANSFECTION and CLONING technologies, sequence and structure analysis algorithms, computer databases, and gene and protein structure function analysis and prediction. Biotechnologies
D012995 Solubility The ability of a substance to be dissolved, i.e. to form a solution with another substance. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed) Solubilities
D013055 Spectrophotometry, Infrared Spectrophotometry in the infrared region, usually for the purpose of chemical analysis through measurement of absorption spectra associated with rotational and vibrational energy levels of molecules. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) IR Spectra,Infrared Spectrophotometry,IR Spectras,Spectra, IR
D016466 CHO Cells CELL LINE derived from the ovary of the Chinese hamster, Cricetulus griseus (CRICETULUS). The species is a favorite for cytogenetic studies because of its small chromosome number. The cell line has provided model systems for the study of genetic alterations in cultured mammalian cells. CHO Cell,Cell, CHO,Cells, CHO
D018929 Cell Culture Techniques Methods for maintaining or growing CELLS in vitro. Cell Culture,Cell Culture Technique,Cell Cultures,Culture Technique, Cell,Culture Techniques, Cell

Related Publications

Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
May 2018, Nature communications,
Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
September 2021, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry,
Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
November 2009, Optics express,
Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
April 2004, Applied spectroscopy,
Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
November 2020, The Science of the total environment,
Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
March 2021, Rheumatology (Oxford, England),
Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
September 2019, Optics letters,
Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
June 2022, Scientific reports,
Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
November 2021, Nature communications,
Florian Capito, and Romas Skudas, and Harald Kolmar, and Christian Hunzinger
June 2023, Optics express,
Copied contents to your clipboard!