Article
Biochemical Research Methods
Rachel Hendricks, Dana Reese, Mark Fedesco, Michael Chinn, Jeffery Zhang, Matt Hutchinson
Summary: The development of manufacturing processes for ADCs poses challenges compared to monoclonal antibodies, requiring additional steps for removing protein aggregates and potentially requiring post-conjugation purification. This paper describes a methodology for developing chromatographic purification steps and its application to three products.
JOURNAL OF CHROMATOGRAPHY A
(2022)
Article
Engineering, Chemical
Sobhana A. Sripada, Driss Elhanafi, Leonard B. Collins, Taufika I. Williams, Marina Y. Linova, John M. Woodley, Cristiana Boi, Stefano Menegatti
Summary: This study presents the development and validation of a novel adsorbent, PichiaGuard, which is functionalized with peptide ligands and targets the whole spectrum of K. phaffii HCPs for protein purification. The PichiaGuard adsorbent has high HCP binding capacity and successfully purified monoclonal antibodies and fragments, showing its potential for modern biopharmaceutical processing.
SEPARATION AND PURIFICATION TECHNOLOGY
(2023)
Article
Biochemical Research Methods
Rongrong Xu, Li Lu, Lingjue Sun, Xiao Liu, Yutian Lei, Shengfeng Huang, Hao Huang, Jacques Crommen, Hai Han, Qiqin Wang, Zhengjin Jiang
Summary: A novel histidine-tagged cyclic peptide monolithic material with excellent affinity and capacity has been developed for the production and purification of monoclonal antibodies. This material not only demonstrates outstanding antibody-specific selectivity, but also effectively prevents aggregation and denaturation of enriched antibodies.
JOURNAL OF CHROMATOGRAPHY A
(2021)
Article
Biotechnology & Applied Microbiology
Roxana Roshankhah, Guoqiang Chen, Yating Xu, Nikhila Butani, Yves Durocher, Robert Pelton, Raja Ghosh
Summary: This study examines the purification of Trastuzumab using cation-exchange z(2) laterally-fed membrane chromatography (z(2)LFMC). The results show that the z(2)LFMC process is comparable to protein A affinity chromatography in terms of purity and recovery, but it is faster and does not require acidic conditions. The productivity of Trastuzumab with the z(2)LFMC process is more than 3 times higher than with column-based purification. Therefore, z(2)LFMC could be a cheaper and more efficient alternative to protein A affinity chromatography.
BIOCHEMICAL ENGINEERING JOURNAL
(2022)
Article
Biotechnology & Applied Microbiology
Ryan R. Davis, Fletcher Suber, Ian Heller, Boxuan Yang, Juan Martinez
Summary: Improving productivity and decreasing costs for biotherapeutic agents is a key driver in the biologics manufacturing industry. Advances in upstream processes have outpaced downstream operations, particularly in monoclonal antibody purification. Utilizing cellulose nanofibers with a protein A ligand in continuous chromatography can greatly increase productivity and reduce costs.
JOURNAL OF BIOTECHNOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
David Saleh, Rudger Hess, Michelle Ahlers-Hesse, Nicole Beckert, Markus Schoenberger, Federico Rischawy, Gang Wang, Joschka Bauer, Michaela Blech, Simon Kluters, Joey Studts, Juergen Hubbuch
Summary: This study investigates the impact of amino acid substitutions in the complementarity-determining region (CDR) of a full-length IgG1 mAb on elution behavior in preparative cation exchange chromatography. The results reveal that single amino acid substitutions can increase charge heterogeneity of the product, which may affect lead optimization during antibody drug discovery.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)
Article
Biochemical Research Methods
Krystian Baran, Patrycja Zimoch, Artur Stanczak, Wojciech Piatkowski, Dorota Antos
Summary: A procedure for adjusting the charge variants of monoclonal antibodies using ion exchange chromatography has been developed, utilizing band splitting phenomenon to separate protein load into enriched or depleted fractions. Cation exchange chromatography (CEX) and anion exchange chromatography (AEX) separations were examined, with the separations based on acidic, main, and basic variants. pH gradient separations were simplified into two stages: isocratic loading and washing, followed by rapid pH change for desorbing remains of protein load. Development of a dynamic model to predict process performance was also discussed.
JOURNAL OF CHROMATOGRAPHY A
(2021)
Article
Biochemical Research Methods
Masakazu Fukuda, Melissa A. Graewert, Cy M. Jeffries, Dmitri I. Svergun, Tadao Yamazaki, Akiko Koga, Yuji Yamanaka
Summary: This study investigates the acidic variants of therapeutic monoclonal antibodies (mAbs) using a refolding approach. The results showed that these acidic variants are conformational variants with minor and localized conformational changes.
ANALYTICAL BIOCHEMISTRY
(2023)
Article
Biochemical Research Methods
Urh Cernigoj, Blaz Nemec, Ales Strancar
Summary: The rise of biosimilar monoclonal antibodies has sparked renewed interest in the composition and separation of monoclonal antibody (mAb) charge variants. Sample displacement chromatography (SDC) has shown potential in enhancing efficiency and productivity in separating mAb charge variants, especially when used in combination with weak cation exchange chromatographic columns. The efficiency of the SDC process is flow rate independent, and at optimal conditions, it can achieve a productivity that is three times higher than the standard bind-elute separation method.
Article
Biochemical Research Methods
Johannes Winderl, Eric Neumann, Jurgen Hubbuch
Summary: Fiber-based CEX adsorbents show comparable performance to resins in removing mAb aggregates, with higher productivity and lower buffer consumption observed in controlled grafted fibers. The use of such adsorbents for mAb aggregate removal is feasible with performance on par with existing resin materials.
JOURNAL OF CHROMATOGRAPHY A
(2021)
Article
Microbiology
Haiqiao Bian, Chong Yu, Yanwu Wei, Li Feng, Changming Liu, Liping Huang
Summary: The study successfully purified PCV2 virus strains using affinity chromatography, resulting in high purity and no significant changes in antigenic characteristics after purification. It was found that PCV2 viruses can be successfully purified, but the yield and purity may vary among different subtypes, and the purified virus can be used for virus structure analysis and viral adsorption and invasion research.
Article
Biophysics
Atsushi Hirano, Shogo Kanoh, Kentaro Shiraki, Momoyo Wada, Masahiro Kitamura, Katsuya Kato
Summary: This study discovered that phosphate-modified zirconia nanoparticles can selectively adsorb immunoglobulin G (IgG) antibodies with high capacity while maintaining their molecular conformation. Compared to traditional Protein A ligands, zirconia nanoparticles have higher affinity for human IgG antibodies, especially IgG3. Due to their chemical and mechanical stability, zirconia nanoparticles can be used in batch methods and chromatography as an alternative to Protein A chromatography for the purification of antibody drugs.
COLLOIDS AND SURFACES B-BIOINTERFACES
(2023)
Article
Chemistry, Multidisciplinary
Jonas Schilz, Charlotte Clement, Franziska Greiner, Arne Skerra
Summary: A novel strategy for the efficient and gentle affinity purification of PASylated proteins is reported, which allows direct one-step purification from cell extracts or culture supernatants while avoiding harsh elution conditions. This method can improve the efficiency of biological drug development.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Biochemical Research Methods
Nikhil Kateja, Anamika Tiwari, Garima Thakur, Anurag S. Rathore
Summary: A continuous mAb platform was developed with both complete continuous (CC) and periodic continuous (PC) modes of operation, showing that CC mode had higher specific productivity and lower consumable utilization, while PC mode was operationally simpler and had lower facility costs due to reduced auxiliary equipment. Both modes offered viable options for continuous processing of mAbs and achieved target critical quality attribute profiles of the final drug product within 24 hours of operation.
BIOTECHNOLOGY JOURNAL
(2021)
Article
Engineering, Chemical
Tomas Molnar, Maria Bartosova, Monika Antosova, Ludovit Skultety, Milan Polakovic
Summary: This study successfully purified rhEPO using a three-step chromatographic procedure, with a purification factor of about 40. The process involved direct chromatographic separation of the protein from the post-harvest medium, with the removal of over 99% of impurity proteins.
SEPARATION AND PURIFICATION TECHNOLOGY
(2021)
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)