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)
Review
Biotechnology & Applied Microbiology
Jia-Wei Liu, Yu-Huan Yang, Nan Wu, Ji-Fu Wei
Summary: This review primarily introduces the current status and approval process of biosimilar monoclonal antibodies in China, analyzes the patent situation in this field, and emphasizes the importance of pharmaceutical companies protecting their intellectual property rights.
Article
Biochemistry & Molecular Biology
Bastiaan Laurens Duivelshof, Thomas Bouvarel, Sebastian Pirner, Vincent Larraillet, Alexander Knaupp, Hans Koll, Valentina D'Atri, Davy Guillarme
Summary: Charge heterogeneity among therapeutic monoclonal antibodies is an important quality attribute that requires careful characterization. The use of solvent-enriched mobile phase composition can improve selectivity for the analysis of various antibody products.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Biochemical Research Methods
Szabolcs Fekete, Amarande Murisier, Matthew Lauber, Davy Guillarme
Summary: This article presents a practical solution to improve the linearity of effluent pH response in pH gradient cation exchange chromatography (CEX) by converting pH to mobile phase volume fraction and generating an inverse response function in the time domain. By setting the inverse function as the gradient program, an ideally linear pH response can be achieved.
JOURNAL OF CHROMATOGRAPHY A
(2021)
Review
Biochemistry & Molecular Biology
Dennis Y. Gout, Lotte S. Groen, Marjolein van Egmond
Summary: Monoclonal antibody (mAb) therapy has been successful in treating lymphomas and leukemias, but its efficacy is reduced in solid tumors due to an immune-suppressive tumor micro-environment. Antibody-cytokine fusion proteins, known as immunocytokines, provide a solution by acting as a local delivery platform and increasing the half-life of cytokines. These fusion proteins can activate cytotoxic immune cells and eliminate tumor cells. Current research focuses on exploring different antibody formats and cytokine payloads for the development of immunocytokines.
CELLULAR AND MOLECULAR LIFE SCIENCES
(2022)
Article
Oncology
Matthew Hummel, Tjerk Bosje, Andrew Shaw, Mark Shiyao Liu, Abhijit Barve, Mudgal Kothekar, Mark A. Socinski, Cornelius F. Waller
Summary: This study demonstrated the bioequivalence of MYL-1402O with EU and US-reference bevacizumab, as well as between EU-reference and US-reference bevacizumab. Pharmacokinetic parameters and safety profiles of MYL-1402O were found to be similar to the reference bevacizumab drugs, with no new significant safety issues emerging.
JOURNAL OF CANCER RESEARCH AND CLINICAL ONCOLOGY
(2022)
Article
Pharmacology & Pharmacy
Yuwei Lu, Chenguang Wang, Bowen Jiang, Changquan Calvin Sun, Stephen W. Hoag
Summary: Oral delivery route provides better patient compliance for biologics targeting gastrointestinal tract. Storage relative humidity can induce conformational changes and aggregation of immunoglobulin G, while commonly used compaction pressures may reduce its binding activity.
INTERNATIONAL JOURNAL OF PHARMACEUTICS
(2021)
Article
Biochemical Research Methods
Izabela Poplewska, Patrycja Zimoch, Dorota Antos
Summary: This study demonstrates the occurrence of pH excursion in pH gradient elution of monoclonal antibodies (mAbs) on strong cation exchange resins under high overloading conditions. The presence of the proteins affects the pH profiles in both liquid and adsorbed phases, and this effect is enhanced with higher initial pH of the binding buffer and protein concentration. The observed pH transitions for different mAbs on the resins predominantly depend on their adsorption affinity.
JOURNAL OF CHROMATOGRAPHY A
(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)
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)
Article
Chemistry, Analytical
Baubek Spanov, Oladapo Olaleye, Tomes Mesurado, Natalia Govorukhina, Alois Jungbauer, Nico C. van de Merbel, Nico Lingg, Rainer Bischoff
Summary: The study found that Pertuzumab undergoes changes in charge heterogeneity, primarily due to deamidation in the Fc domain and N-terminal pyroglutamate formation in the heavy chain. Peptide mapping analysis also showed that clinical samples had 2-3% deamidation in the heavy chain CDR2, 20-25% deamidation in the Fc domain, and 10-15% N-terminal pyroglutamate formation in the heavy chain.
ANALYTICAL CHEMISTRY
(2023)
Article
Immunology
Long Liu, Lu Qi, Chunpu Lei, Yu Wang, Wei Zhang, Ying Liu, Pu Li, Haihong Bai, Yan Li, Yinjuan Li, Ju Liu, Liangzhi Xie, Xinghe Wang
Summary: This study compared the pharmacokinetics, safety, and immunogenicity of the adalimumab biosimilar SCT630 with its reference drug. The results showed that SCT630 was bioequivalent to adalimumab in terms of pharmacokinetic parameters, and had similar safety and immunogenicity profiles.
INTERNATIONAL IMMUNOPHARMACOLOGY
(2022)
Article
Chemistry, Medicinal
Dong Gao, Lei Nie, Junjie Yuan, Feng Hu, Zhenhua Wu, Qunhai Lin, Haibin Wang
Summary: The biosimilar drug HS016 and the reference product adalimumab (Humira) were characterized and compared using physical chemistry and functional similarity tests. While slight differences were observed in certain characteristics, the primary and higher order structures, physicochemical properties, and biological activity of HS016 and adalimumab were found to be similar.
JOURNAL OF PHARMACEUTICAL SCIENCES
(2022)
Article
Biochemical Research Methods
Md Abdullah-Al-Kamran Khan, Rafeed Rahman Turjya, Abul Bashar Mir Md Khademul Islam
Summary: This study proposed a systematic in silico approach to design new variants of Adalimumab mAb with improved binding properties, resulting in one significantly improved variant out of 143 models. Six important residues were identified, and suitable mutations were applied to design significant variants, offering new insights for structure-based affinity improvements of monoclonal antibodies.
JOURNAL OF MOLECULAR GRAPHICS & MODELLING
(2021)
Article
Biochemistry & Molecular Biology
Hyejin Kim, Geul Bang, Ye Eun Park, Moonhee Park, Jung Hoon Choi, Myung Jin Oh, Hyun Joo An, Jong Shin Yoo, Youngja Hwang Park, Jin Young Kim, Heeyoun Hwang
Summary: Characterization of therapeutic monoclonal antibodies (mAbs) is challenging due to their heterogeneity caused by post-translational modifications (PTMs). In this study, we used high-resolution tandem mass spectrometry and ultra-high-performance liquid chromatography to characterize and compare biologics and biosimilar of infliximab. By comparing N- and O-glycopeptides profiles, we identified differences between the biologics and biosimilar, including novel N-glycosylation sites at the light chain and an O-glycopeptide at the heavy chain in the biosimilar only. Site-specific glycopeptide analysis is a reliable technique for evaluating therapeutic mAbs and complex glycoprotein products.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2022)
Article
Biochemical Research Methods
Leo A. A. Jakob, Tomas Mesurado, Alois Jungbauer, Nico Lingg
Summary: The CASPON enzyme is an interesting enzyme for fusion protein processing due to its generation of an authentic N-terminus. However, the high cysteine content in CASPON may lead to aggregation through disulfide-bond formation, which can negatively impact enzymatic activity. Different multimerization states of CASPON were isolated and analyzed to determine their propensity for multimerization and enzymatic activity. The results showed that the dimer state was the most stable and active, with 50% higher enzymatic activity than the tetramer state. Multimerization was mainly mediated by cysteine, and attractive protein-protein interactions were observed in the presence of ammonium sulfate and on a chromatographic stationary phase. These findings provide insights into protein purification and highlight the importance of controlling multimerization to maintain protein activity.
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
(2023)
Article
Chemistry, Analytical
Baubek Spanov, Oladapo Olaleye, Tomes Mesurado, Natalia Govorukhina, Alois Jungbauer, Nico C. van de Merbel, Nico Lingg, Rainer Bischoff
Summary: The study found that Pertuzumab undergoes changes in charge heterogeneity, primarily due to deamidation in the Fc domain and N-terminal pyroglutamate formation in the heavy chain. Peptide mapping analysis also showed that clinical samples had 2-3% deamidation in the heavy chain CDR2, 20-25% deamidation in the Fc domain, and 10-15% N-terminal pyroglutamate formation in the heavy chain.
ANALYTICAL CHEMISTRY
(2023)
Article
Biotechnology & Applied Microbiology
Magdalena Pappenreiter, Benjamin Bayer, Marijan Logarusic, Bernhard Sissolak, Alois Jungbauer
Summary: The study investigated the individual impacts of mild and rigorous intra-experimental parameter shifts on the culture performance and cell metabolism of an antibody producing CHO cell line. The results showed significant changes in cell growth, metabolism, and productivity due to intra-experimental parameter shifts. However, if the conditions within an experiment are changed, rigorous parameter shifts should be avoided, as they can lead to irreversible changes in cell behavior, as observed in media blending experiments.
BIOCHEMICAL ENGINEERING JOURNAL
(2023)
Article
Biotechnology & Applied Microbiology
Viktoria Mayer, Anna-Carina Frank, Shirin Preinsperger, Patrick Csar, Petra Steppert, Alois Jungbauer, Patricia Pereira Aguilar
Summary: Host cell DNA, especially chromatin, is an important impurity in downstream processing of enveloped viruses. Endonuclease treatment, using salt-tolerant endonucleases, combined with flow-through chromatography using core-shell particles, showed high purity and purification efficiency in the processing of recombinant measles virus. This strategy has the potential to be applied in other viral platforms.
BIOTECHNOLOGY PROGRESS
(2023)
Article
Biotechnology & Applied Microbiology
Magdalena Pappenreiter, Hubert Schwarz, Bernhard Sissolak, Alois Jungbauer, Veronique Chotteau
Summary: This study tested different process set ups and conditions in low shear TFF systems using magnetic levitation pumps in small-scale perfusion bioreactors. The authors validated a novel concept of applying reverse flow across the hollow fiber using two magnetically levitating pumps. The results showed that product sieving could be improved by 30% with the application of dynamic recirculation flow, and minimal product retention was achieved by reversing the flow of two alternating pumps with short cycle times. Additionally, a correlation was found between the passage of high molecular weight species and the flow direction, as well as the degree of product sieving.
JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY
(2023)
Review
Biotechnology & Applied Microbiology
Astrid Duerauer, Alois Jungbauer, Theresa Scharl
Summary: The biopharmaceutical industry is still running in batch mode due to heavy regulation, but there are initiatives and technologies being developed to move towards automated and autonomous bioprocessing. These include Process Analytical Technologies, Quality by Design, and Continuous Integrated Manufacturing. The ultimate goal is to achieve real-time release and establish a highly predictive and robust biomanufacturing system.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Article
Biotechnology & Applied Microbiology
Gabriele Recanati, Magdalena Pappenreiter, Christoph Gstoettner, Patrick Scheidl, Elena Dominguez Vega, Bernhard Sissolak, Alois Jungbauer
Summary: Continuous precipitation coupled with continuous tangential flow filtration is an economical method for capturing recombinant antibodies from cell culture supernatant. By adopting tubular reactors to replace surge tanks, a continuous harvest and mass flow of product can be achieved with a narrow residence time distribution. The combination of CaCl2 precipitation and PEG capture method allows for efficient removal of host-cell DNA and retention of glycosylation profile.
ENGINEERING IN LIFE SCIENCES
(2023)
Article
Biochemical Research Methods
Markus Mozgovicz, Andreas Fischer, Cecile Brocard, Alois Jungbauer, Nico Lingg
Summary: Immobilized metal affinity chromatography (IMAC) is a powerful technique for capturing and purifying biopharmaceuticals. Protein recovery can be compromised due to spreading and unfolding of the protein, leading to fouling of the stationary phase. This study investigates the kinetics of irreversible adsorption of a protease on an IMAC resin and suggests that L-arginine sulfate can decelerate the irreversible adsorption.
JOURNAL OF CHROMATOGRAPHY A
(2023)
Article
Biochemistry & Molecular Biology
Nico Lingg, Markus Mozgovicz, Theresa Schaufler, Alexander Prokup, Amy Carruthers, David Boerema, Laura Keigher, Nathan Brinkman, Alois Jungbauer
Summary: Currently, IVIG is manufactured in batch through cold ethanol precipitation, which leads to a slow process with high space requirements. We have developed a continuous method using a tubular reactor to replace the batch precipitation method. This reactor, filled with static mixers for efficient cooling, allows for complete precipitation with the same yield and purity even with a residence time of only one minute. A tubular reactor with specific dimensions can process 4000 L of plasma per day, providing an opportunity to reduce the plant footprint or intensify existing manufacturing plants.
PROCESS BIOCHEMISTRY
(2023)
Article
Engineering, Chemical
Mafalda S. Santos, Christian Schuster, Harald Rennhofer, Helga C. Lichtenegger, Herwig Peterlik, Tim Causon, Alois Jungbauer
Summary: Efficient recovery of building-block chemicals, such as biobased itaconic acid, produced by microbial processes can be achieved using ultrathin membranes. The composition of the membranes was optimized to improve the separation performance, and membranes with high selectivity for itaconic acid were successfully demonstrated in experiments with synthetic solutions and culture supernatant.
SEPARATION AND PURIFICATION TECHNOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Hannah Engelmaier, Christoph Dattenboeck, Katerina Petrushevska-Seebach, Alois Jungbauer, Nikolaus Hammerschmidt
Summary: The study presents an integrated, continuous process for the capture of recombinant blood coagulation factors. The developed process consists of three continuous unit operations and has demonstrated successful continuous capture of the factors over a 24-hour duration.
JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY
(2023)