Article
Chemistry, Medicinal
Ryan Knihtila, Yuanli Song, Letha Chemmalil, Julia Ding, Nesredin Mussa, Zheng Jian Li
Summary: Size Exclusion Chromatography (SEC) is widely used in assessing aggregate content in bio-pharmaceutical drugs like monoclonal antibodies (mAbs), with electrostatic interactions between protein analytes and SEC column resin needing to be minimized. Increasing mobile phase salt concentration can decrease electrostatic interactions but may increase protein hydrophobicity and interactions with the stationary phase. The optimized SEC method conditions were found to be robust and suitable for product release and stability studies, with arginine identified as an essential additive to minimize undesirable hydrophobic interactions.
JOURNAL OF PHARMACEUTICAL SCIENCES
(2021)
Article
Chemistry, Analytical
Gao-Yuan Liu, Zhengqi Zhang, Yuetian Yan, Shunhai Wang, Ning Li
Summary: In this study, a novel covalent cross-link formed via an imine bond between lysine and serine residues was discovered in therapeutic monoclonal antibodies (mAbs) using a multipronged MS-based approach. This Ser-Lys cross-link contributes to the formation of covalent high-molecular-weight species in mAb products.
ANALYTICAL CHEMISTRY
(2023)
Article
Chemistry, Analytical
Prity Kumari, Marianne Saldanha, Ratnesh Jain, Prajakta Dandekar
Summary: Addition of sugar osmolytes and formulation excipients can effectively reduce monoclonal antibody aggregation during cell culture, improving downstream processing and process yield.
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
(2023)
Article
Chemistry, Analytical
Thuy Tran, Erik Martinsson, Sergio Vargas, Ingemar Lundstrom, Carl-Fredrik Mandenius, Daniel Aili
Summary: The production of therapeutic monoclonal antibodies is a complex process that requires extensive analytical characterization. Aggregation of antibodies is common and problematic, as it can affect the product quality and immunogenicity. Rapid detection of aggregates is challenging with current analytical techniques.
ANALYTICAL CHEMISTRY
(2022)
Article
Biotechnology & Applied Microbiology
Young Hoon Oh, Matthew L. Becker, Kerri M. Mendola, Leila H. Choe, Lie Min, Kelvin H. H. Lee, Yinges Yigzaw, Alexander Seay, Jerome Bill, Xuanwen Li, David J. Roush, Steven M. Cramer, Stefano Menegatti, Abraham M. Lenhoff
Summary: In the production of biopharmaceuticals, the presence of residual host-cell proteins (HCPs) is a critical quality concern. Measurements on seven monoclonal antibody (mAb) processes revealed that HCP aggregates, not necessarily derived from chromatin, are important factors in the persistence of HCPs. These aggregates can also interfere with accurate detection of HCPs using proteomics methods. The study also highlights the difficulty in removing certain HCPs due to their functional complementarity to the biopharmaceutical product.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Article
Multidisciplinary Sciences
Nicole L. McIntosh, Geoffrey Y. Berguig, Omair A. Karim, Christa L. Cortesio, Rolando De Angelis, Ayesha A. Khan, Daniel Gold, John A. Maga, Vikas S. Bhat
Summary: Size exclusion chromatography coupled to multiangle light scattering is a straightforward method for comprehensive characterization of AAV capsids, providing detailed information with high accuracy and precision. By using new equations and intrinsic properties, multiple physical attributes of AAV can be quantified in a short time, making this method a powerful tool for future gene therapy programs.
SCIENTIFIC REPORTS
(2021)
Article
Biochemical Research Methods
Dean Lee, Yucheng Zhu, Byeong Chae, Elizabeth J. Tocce
Summary: This study investigated the experimental conditions required to obtain the molecular weight distribution of ultrahigh molecular weight poly(ethylene oxide) (PEO) using SEC hyphenated with MALS and RI. Different fitting approaches resulted in significant differences in molecular weight, with a proposal to use specific angle signals for molecular weight derivation.
JOURNAL OF CHROMATOGRAPHY A
(2021)
Article
Chemistry, Analytical
Kai-Wei Wu, Tse-Hong Chen, Teng-Chieh Yang, Shao-Chun Wang, Mohammed Shameem, Kenneth S. Graham
Summary: Size exclusion chromatography (SEC) is a foundational analytical method for assessing the purity of biological molecules. Monitoring the efficiency and continuity of the chromatographic column is crucial for obtaining accurate and reproducible data that meet regulatory standards. In this study, 19 SEC columns were evaluated using an in-house monoclonal antibody. System suitability parameters were used to monitor the performance of the SEC assay, revealing that certain factors increased with injection number while others decreased. After 1000 injections, specific factors showed significant changes in comparison to their starting values.
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
(2023)
Article
Medicine, Research & Experimental
Shravan Sreenivasan, Anurag S. Rathore
Summary: This study investigated the aggregation of mAb in the presence of Fe2+ and H2O2 in saline and in vitro models. The results showed that the combined presence of Fe2+ and H2O2 increased mAb aggregation and altered protein structure and hydrophobicity. Increasing the time, temperature, and Fe2+ and H2O2 concentration further enhanced mAb aggregation. Additionally, mAb samples with copper and cobalt chlorides also resulted in degradation.
MOLECULAR PHARMACEUTICS
(2023)
Article
Chemistry, Medicinal
Yukie Yoshii, Hiroki Matsumoto, Chika Igarashi, Tomoko Tachibana, Fukiko Hihara, Mitsuhiro Shinada, Atsuo Waki, Sei Yoshida, Kenichiro Naito, Kimiteru Ito, Tatsuya Higashi, Hiroaki Kurihara, Makoto Ueno
Summary: Understanding the physicochemical properties of antibody-drug conjugates is crucial for assessing their quality and monitoring their storage. In this study, the larger size variants contained in the antibody-chelator conjugate were characterized, and a chromatographic method was developed to remove these size variants. The study demonstrated the stability of the product after the removal of size variants and suggested that the developed methods can ensure the quality of PCTA-NCAB001 and other antibody-drug conjugates.
Article
Materials Science, Multidisciplinary
Ryan A. Fair, Renxuan Xie, Youngmin Lee, Ralph H. Colby, Enrique D. Gomez
Summary: Molecular weight is a critical parameter for polymers, especially conjugated polymers, often measured using gel permeation chromatography (GPC). Due to mismatch in chain flexibilities between samples and standards, as well as poor solubility in the mobile phase, inaccuracies in GPC measurements of conjugated polymers are common. By applying a universal calibration method and in-line concentration measurements, accurate results for polymers of various stiffnesses can be obtained, with the accuracy verified using absolute molecular weights from static light scattering. Measuring the refractive index increment is crucial for confirming full recovery of the polymer and ensuring accurate values of absolute molecular weight from GPC.
ACS APPLIED POLYMER MATERIALS
(2021)
Article
Pharmacology & Pharmacy
Haresh T. More, Dilbir S. Bindra, Alexandra Zumba, Kaimeng Zhou, Thiago Carvalho, Rao Mantri
Summary: In this study, it was found that therapeutic proteins, such as monoclonal antibodies (mAbs), are exposed to ambient light conditions during manufacturing and handling processes. The RT/RL stability studies are generally conducted to determine the allowable exposure time. However, an unexpected increase in protein aggregation was observed during a formal RT/RL study due to differences in the setup of the stability chamber. This study emphasizes the importance of understanding UV levels and using representative light conditions for accurate stability assessment of biologic drug products.
EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS
(2023)
Article
Plant Sciences
Sukhita Sathitnaitham, Anongpat Suttangkakul, Passorn Wonnapinij, Simon J. McQueen-Mason, Supachai Vuttipongchaikij
Summary: Cell walls are dynamic, multi-component materials that are crucial in plant biology, with their properties modulated by wall-related enzymes. A new method allows simultaneous examination of specific cell wall matrix components, providing insight into direct and indirect consequences of cell wall manipulations. By utilizing this method, researchers can gain a deeper understanding of wall composition, its dynamism, and plasticity, further advancing knowledge of cell wall biology.
Article
Pharmacology & Pharmacy
Ashutosh Sharma, Jason Beirne, Dikshitkumar Khamar, Ciaran Maguire, Ambrose Hayden, Helen Hughes
Summary: Biopharmaceuticals are complex and unstable therapeutic molecules, and their safety and quality need to be ensured. This study evaluated the application of MADLS to measure particle size, concentration, and aggregation in different protein modalities. The results showed that MADLS can be used as a 3-in-1 approach to assess particle size, concentration, and aggregation, reducing the number of required samples and experiments.
Article
Biochemical Research Methods
Pauline Legrand, Oumar Dembele, Helena Alamil, Catherine Lamoureux, Nathalie Mignet, Pascal Houze, Rabah Gahoual
Summary: The translation text describes an analytical strategy that enables the structural identification and quantification of monoclonal antibodies in potentially counterfeit drugs. By using capillary zone electrophoresis analysis, size exclusion chromatography coupled to multi-angle light scattering, and liquid chromatography tandem mass spectrometry, information on the nature and structure of mAbs is obtained.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2022)