Article
Biochemical Research Methods
Meng Li, Xueyu Zhao, Danfeng Shen, Gang Wu, Wenbo Wang, Chuanfei Yu, John Sausen, Hanmei Xu, Lan Wang
Summary: This article describes the method of using capillary electrophoresis-mass spectrometry (CE-MS) to characterize the critical quality attributes (CQAs) of recombinant monoclonal antibody therapeutics. By combining the CZE-MS and cIEF-MS methods with the traditional CE-SDS analysis workflow, a comprehensive profile of monoclonal antibody size variants can be obtained.
JOURNAL OF CHROMATOGRAPHY A
(2022)
Article
Biochemical Research Methods
Jason Candreva, Abbie L. Esterman, Derek Ge, Pritesh Patel, Shannon C. Flagg, Tapan K. Das, Xue Li
Summary: The clinical benefits of combination treatments with therapeutic monoclonal antibodies (mAbs) have become apparent. In this study, a novel methodology for exploring charge variants of mAb mixtures using multiple detection techniques simultaneously is reported. This approach allows accurate quantification of charge variants in mixtures with ratio differences of up to 40-fold.
Article
Chemistry, Analytical
Cecile Tardif, Emmanuel Jaccoulet, Jean-Francois Bellec, Yannick Surroca, Laurence Talbot, Myriam Taverna, Claire Smadja
Summary: Monoclonal antibodies are widely used in cancer therapy and require reliable characterization methods. The combination of image capillary isoelectric focusing and chemometric approaches, such as PCA and PLS-DA, has been investigated for identifying and predicting therapeutic monoclonal antibodies. The analysis of four commercialized mAbs using icIEF-PCA resulted in clear clustering and successful prediction. This approach proves to be a reliable method to ensure the quality of compounded therapeutic monoclonal antibodies before patient administration.
Review
Chemistry, Multidisciplinary
Tian Xu, Liangliang Sun
Summary: Capillary isoelectric focusing (cIEF) combined with electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) is an ideal method for characterizing proteoforms based on their isoelectric points, but faces technical challenges that impede widespread adoption.
FRONTIERS IN CHEMISTRY
(2021)
Review
Chemistry, Analytical
Filip Dusa, Anna Kubesova, Jiri Salplachta, Dana Moravcova
Summary: Capillary isoelectric focusing (cIEF) is a powerful separation technique that requires the repeatable formation of a pH gradient and the use of internal standards to track its profile. This review provides an overview of available isoelectric point markers and their syntheses, as well as recent applications of cIEF in analyzing complex larger structures.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2023)
Article
Chemistry, Analytical
Alexander B. Schwahn, Julia Baek, Shanhua Lin, Christopher A. Pohl, Ken Cook
Summary: Characterization and quality control of biotherapeutic proteins often require multiple orthogonal separation techniques, leading to tedious method development. This study demonstrates the use of a linear controlled pH gradient system and universal buffer solutions to improve efficiency and minimize manual intervention in method development and analysis.
ANALYTICAL CHEMISTRY
(2022)
Article
Biochemical Research Methods
Teresa Kwok, She Lin Chan, Matthew Courtney, Mike Zhou, Tiemin Huang, Tao Bo, Victor Li, Tong Chen
Summary: Recombinant monoclonal antibodies have driven the growth of commercial biotherapeutics. In this study, a nano-ESI apparatus was constructed to combine icIEF with HRMS, allowing for a comprehensive understanding of protein charge heterogeneity. The system demonstrated excellent sensitivity and repeatability, making it a valuable tool for assessing the structural modifications impacting protein charge heterogeneity.
ANALYTICAL BIOCHEMISTRY
(2023)
Article
Chemistry, Analytical
Filip Dusa, Dana Moravcova, Karel Slais
Summary: We present a set of novel low-molecular-mass compounds with ampholytic properties that can be used as markers of isoelectric point in different isoelectric focusing formats. These compounds have excellent focusing properties and stability, making them suitable for various isoelectric focusing methods.
ANALYTICA CHIMICA ACTA
(2022)
Article
Pharmacology & Pharmacy
Gang Wu, Chuanfei Yu, Wenbo Wang, Jialiang Du, Gangling Xu, Zhihao Fu, Lan Wang
Summary: This study developed a fully automated capillary isoelectric focusing-mass spectrometry (cIEF-MS) workflow for charge variant analysis of bispecific antibody molecules. The workflow showed clear separation and accurate identification of charge variants in the tested BisAb example. The method exhibited high resolution, system sensitivity, robustness, and reproducibility, making it a powerful assay for quality assessment of recombinant protein therapeutics.
CURRENT PHARMACEUTICAL ANALYSIS
(2022)
Article
Biochemical Research Methods
Jun-Jie Yuan, Dong Gao, Feng Hu, Yang Shi, Zhen-Hua Wu, Chuan-Qin Hu, Xiao-Dong Huang, Wei-Jie Fang, Hai-Tao Zhang, Hai-Bin Wang
Summary: This research investigated and characterized charge variants in monoclonal antibodies, identifying the main causes and effects. Through extensive analysis, it was concluded that charge heterogeneity does not impact biological functions. The study suggests that charge variants should be considered effective components, unless there is a significant impact on biological functions.
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
(2021)
Review
Biochemical Research Methods
Gyula Vigh, Bohuslav Gas
Summary: In 1961, Svensson first described the principle of isoelectric focusing (IEF), which used a natural pH gradient formed by passing current through a sucrose density gradient-stabilized ampholyte mixture in a constant cross-section apparatus to separate ampholytic compounds. When polyacrylamide gel replaced the sucrose density gradient, the spatial and temporal stability of Svensson's pH gradient was lost, leading to a search for an explanation and improvement. It was later suggested that in the modern open-system IEF experiment, a two-stage process occurs, with a rapid first phase of linear pH gradient development followed by a slow second stage where isotachophoretic processes cause the pH gradient to decay as extreme pI CAs move into the electrode vessels.
Review
Chemistry, Analytical
Jiaqi Wu, Will McElroy, Janusz Pawliszyn, Christopher D. Heger
Summary: This article reviews the applications of icIEF technology in the pharmaceutical industry, highlighting the benefits of whole-column imaging detection for capillary isoelectric focusing. It also explores recent research using this technology to support advancements in the therapeutic landscape.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Analytical
Zhijie Wu, Hongxia Wang, Jikang Wu, Yu Huang, Xueqing Zhao, Jennifer B. Nguyen, Michael P. Rosconi, Erica A. Pyles, Haibo Qiu, Ning Li
Summary: Therapeutic antibodies are important drugs used to treat diseases, and their characterization is crucial for drug efficacy and safety. Charge variant analysis using nCE-MS is an alternative method to support the characterization of therapeutic antibodies. In this study, nCE-MS analysis was performed to evaluate the charge variants and impurities in therapeutic antibodies, and high-resolution separation and sensitive detection of low abundance species were achieved. This method has significant implications for biopharmaceutical development and investigations.
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
(2023)
Review
Chemistry, Analytical
Harleen Kaur, Jeff Beckman, Yiting Zhang, Zheng Jian Li, Marton Szigeti, Andras Guttman
Summary: Capillary electrophoresis is a powerful technique for comprehensive physicochemical characterization of therapeutic biomolecules, such as monoclonal antibodies, providing high resolution separation and high sensitivity characterization. Techniques like SDS-CGE, CZE, and iCIEF are increasingly used to assess heterogeneity in mAbs and related therapeutic modalities, with advantages and disadvantages that are critically discussed.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2021)
Article
Chemistry, Analytical
Baubek Spanov, Oladapo Olaleye, Tomes Mesurado, Natalia Govorukhina, Alois Jungbauer, Nico C. van de Merbel, Nico Lingg, Rainer Bischoff
Summary: The ion-exchange profile of pertuzumab was evaluated, revealing that deamidation in the Fc domain and N-terminal pyroglutamate formation in the heavy chain were the main contributors to charge heterogeneity. Peptide mapping analysis of clinical samples showed deamidation rates of 2-3% in the heavy chain CDR2, 20-25% in the Fc domain, and N-terminal pyroglutamate formation rates of 10-15% in the heavy chain. These findings suggest that in vitro stress studies can predict in vivo modifications.
ANALYTICAL CHEMISTRY
(2023)
Correction
Biochemical Research Methods
Yong Wang, Suhua Yu, Rui Li, Jing Wan, Yuheng Wang, Zongli Huo, Chunyong Wu, Li Mi, Songqin Liu
JOURNAL OF CHROMATOGRAPHY A
(2024)
Review
Biochemical Research Methods
Israel D. Souza, Maria Eugenia C. Queiroz
Summary: Organic-silica hybrid monolithic materials have gained significant attention in separation science and sample preparation due to their unique properties. This review summarizes the characteristics, current status, and recent developments of these materials, as well as discusses synthesis methods, characterization techniques, and strategies for improving extraction efficiency in various applications.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Bhagwat Upreti, Bhanu Prasad Vempatapu, Jagdish Kumar, Pankaj K. Kanaujia
Summary: Lubricating oils are essential for the effective performance of internal combustion engines. This study presents a new HPLC method for determining additives in finished lubricating oils and polar substances in used engine oils.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Zongqing Huang, Haoju Hua, Xiuzhen Du, Zipeng Zhen, Wei Zhao, Jun Feng, Ji-an Li
Summary: This study successfully isolated VHHs against the SUMO protein for the first time using biopanning of an immune camelid nanobody library. The VHHs were coupled to agarose resins to form affinity chromatography resins, allowing for one-step purification of various SUMO fusions.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Xueting Qiu, Xuben Hou, Yue Yang, Hao Fang, Fei Cui, Xinying Yang
Summary: In this study, a new method based on electrophoretically mediated microanalysis was developed for the investigation of PTPRO enzymatic activity. The method was successfully used to determine the kinetics of PTPRO and the inhibitory concentration of PTPRO inhibitors. The method exhibited improved accuracy and reduced consumption compared to traditional methods.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Feifei Luo, Mengmeng Hao, Lei Zhang, Yangguo Xie, Wei Hou, Hongya Wang, Zhongli Zhang
Summary: In this study, a comprehensive screening and structure elucidation of 23 nonvolatile organic compounds (NVOCs) in two antibody drugs was conducted. Seven previously unreported compounds were identified. A quantitative method with high sensitivity, accuracy, and precision was developed for evaluating the safety of NVOCs in drug products.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Zuzana Vosahlova, Kveta Kalfkova, Martin Gilar, Jakub Szymarek, Maria Mazurkiewicz-Beldzinska, Sylwia Studzi
Summary: A hydrophilic interaction liquid chromatography method was developed for the separation and identification of oligonucleotide impurities and metabolites, and successfully applied to the analysis of serum samples from Spinraza-treated patients.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Aswanidevi Kongasseri, Thirumalai Madhesan, Sangeetha Krishna Kumar, Sushmitha Pedugu Sivaraman, Suchashrita Mitra, Pitchaiah Kancharlapalli Chinaraga, C. V. S. Brahmmananda Rao, Sivaraman Nagarajan, Prabhakaran Deivasigamani, Akhila Maheswari Mohan
Summary: This study focuses on the sequential separation of trivalent lanthanides using modified C-18 silica-packed supports and reversed-phase HPLC technique. By tuning the column modifiers and separation parameters, the researchers achieved systematic separation of Ln(3+). The results showed that the choice of column modifiers and fine-tuning of the modification procedure were crucial for achieving excellent separation.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Raquel Perez-Robles, Szabolcs Fekete, Robert Kormany, Natalia Navas, Davy Guillarme
Summary: There is a mismatch between the sample diluent and the mobile phase in HILIC analysis, leading to analyte breakthrough and peak distortion. Adding a weak solvent for pre-injection can effectively prevent these issues and improve sensitivity.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Oleg Pokrovskiy, Irina Rostovschikova, Konstantin Ustinovich, Ilya Voronov, Dmitry Kosyakov
Summary: A chromatography-like propagation of water through the material bed was observed during supercritical fluid extraction of aroma plants with high moisture content, which could be significant for modeling the kinetics of supercritical fluid extraction of non-volatile compounds.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Yasar Kemal Recepoglu, Ozguer Arar, Asli Yuksel
Summary: Biosorption using phosphorylated hazelnut shell waste (FHS) is a promising method for lithium removal and recovery. This study investigated the performance of a continuous-flow packed-bed column system using FHS for ultra-low level lithium removal. The results showed that the lihium biosorption capacity of the FHS column was unaffected by the bed height, but decreased with increasing flow rate.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Jian Sun, Zheheng Zheng, Zixiao Jia, Jiabin Wang, Xucong Lin
Summary: In this study, an ionic covalent organic framework (iCOF) surface grafting monolithic sorbent was prepared for in-tube solid-phase microextraction (SPME) of aristolochic acids (AAs) in various samples. The density of quaternary ammonium ions in the iCOF was modulated for enhanced adsorption of AAs. The iCOF surface grafting monolithic sorbent was confirmed by different means. Several operating conditions were optimized to develop the online in-tube SPME-HPLC method, which showed good linearity, low limits of detection, and high recoveries for AAs in serum samples. This study presents a feasible approach for the application of COF-based monolithic sorbents in sample preparation.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Zeng Wen, Ruihan Peng, Die Gao, Jing Lin, Jia Zeng, Zhou Li, Famin Ke, Zhining Xia, Dandan Wang
Summary: A chitosan-alginate sponge with multiple cross-linking networks was developed for adsorption and enrichment of anionic dyes from food samples. The sponge showed better three-dimensional network structure, mechanical behavior, and stability compared to pure chitosan and alginate sponges. The adsorption performance and selectivity of the sponge were systematically studied, and it maintained high adsorption capacity and selectivity for anionic dyes. The sponge also showed satisfactory reusability and was successfully combined with high-performance liquid chromatography for the enrichment and detection of anionic dyes in candy and cocktail samples.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Hairong Zhang, Xiaojuan Jiang, Dandan Zhang, Yuexin Yang, Qiang Xie, Caisheng Wu
Summary: Analysis of in vivo exposure to traditional Chinese medicine using mass spectrometry is helpful for identifying effective ingredients and developing new drugs. This study focused on Forsythia suspensa (Lian Qiao) and used metabolomics technology to explore its in vivo exposure. By comparing the differences before and after administration, a complete set of nontargeted analysis strategies for TCM exposure was established. The study identified 393 Lian Qiao-related chemical components in mice after administration, including prototypes and metabolites. This study provides a reference for research on the active ingredients of Lian Qiao in vivo.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biochemical Research Methods
Oscar Castejon-Musulen, Ricardo Lopez, Ignacio Ontan, Vicente Ferreira
Summary: A quantitative analytical method was developed and validated in this study to determine the concentrations of 81 aroma-relevant wine volatiles. The method utilizes stir bar sorptive extraction and multidimensional gas chromatography with flame ionization detector or mass spectrometer. The method demonstrated good linearity, reproducibility, and recovery, and was successfully applied to analyze 20 wine samples.
JOURNAL OF CHROMATOGRAPHY A
(2024)