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
Amarande Murisier, Bastiaan L. Duivelshof, Szabolcs Fekete, Julien Bourquin, Andrew Schmudlach, Matthew A. Lauber, Jennifer M. Nguyen, Alain Beck, Davy Guillarme, Valentina D'Atri
Summary: This study successfully demonstrated the direct hyphenation of cation exchange chromatography with a compact Time of Flight mass spectrometer for analyzing monoclonal antibodies. By testing various buffers and column dimensions, optimal chromatographic conditions were identified for rapid analysis of charge variants.
JOURNAL OF CHROMATOGRAPHY A
(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)
Article
Biochemical Research Methods
Gaoya Yuan, Xudong Zhang, Yuanyi Zhang, Wanyuan Dong, Yifeng Li
Summary: In this work, a method for continuous separation and collection of acidic and basic charge variants was developed, enabling efficient generation of these variants with high purity.
PROTEIN EXPRESSION AND PURIFICATION
(2023)
Article
Pharmacology & Pharmacy
Busra Gurel, Melike Berksoz, Eda Capkin, Ayhan Parlar, Meltem Corbacioglu Pala, Aylin Ozkan, Yilmaz Capan, Duygu Emine Daglikoca, Meral Yuce
Summary: Avastin (R) is a recombinant monoclonal antibody used in cancer treatment. The biosimilar candidate SIMAB054 developed in this study showed different charge variant profile compared to the innovator product. The structural and functional analysis revealed that they share common glycoforms and post-translational modifications, but with different ratios in some samples. The differences mainly originated from the presence of extra C-terminal lysine residues.
Article
Immunology
Dag Nyman, Marika Nordberg, Clara Nyberg, Susanne Olausson, Nellie Carlstromer Berthen, Sten-Anders Carlsson
Summary: Lyme disease, caused by Borrelia burgdorferi, is a tick-borne infection that continues to spread in temperate regions. The clinical diagnosis of Borrelia infection is challenging and current laboratory tests have limitations. A novel assay for quantitating IgG1-subclass antibodies showed comparable performance to a commercial reference assay. However, in high-endemic areas, antibody tests cannot accurately diagnose active cases.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2023)
Article
Biochemical Research Methods
Liesa Verscheure, An Cerdobbel, Pat Sandra, Frederic Lynen, Koen Sandra
Summary: This study reported a fully automated method for characterizing monoclonal antibody charge variants, which significantly reduces operation time and sample loss, and improves information gathering efficiency. The method can effectively reveal major and minor charge variants, as well as post-translational modifications and modification sites.
JOURNAL OF CHROMATOGRAPHY A
(2021)
Article
Chemistry, Analytical
Aran Kim, Kanghyuk Choi
Summary: This study examines the effect of amines as additives for separating neighboring lanthanides and efficiently producing carrier-free Lu-177. Methylamine is found to be the most effective additive for separation and purification, allowing for successful separation of Lu and Yb.
JOURNAL OF RADIOANALYTICAL AND NUCLEAR CHEMISTRY
(2022)
Article
Biochemical Research Methods
Sunil Kumar, Vineela Peruri, Anurag S. Rathore
Summary: In this study, an online volatile-salt-containing two-dimensional liquid chromatography (2D-LC) method coupled with mass spectrometry (MS) was used to characterize the charge heterogeneity of monoclonal antibodies (mAbs). The 2D-LC method successfully separated and identified more mAb variants, including minor variations in charge distribution or conformation that could not be detected by single analytical methods alone.
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
(2023)
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
Medicine, General & Internal
Juan Carlos Cordova, Sheng Sun, Jeffrey Bos, Srinath Thirumalairajan, Sanjeevani Ghone, Miyako Hirai, Ricarda A. Busse, Julia S. v. der Hardt, Ian Schwartz, Jieyu Zhou
Summary: Membrane chromatography, specifically utilizing Sartobind(R) S and Phenyl membranes, is used to remove aggregates and excess payload in ADC downstream processing. This approach effectively improves the purity of ADCs by clearing unwanted species and increasing homogeneity of drug-to-antibody ratio (DAR).
JOURNAL OF CLINICAL MEDICINE
(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
Heikki Saari, Reetta Pusa, Heli Marttila, Marjo Yliperttula, Saara Laitinen
Summary: The traditional methods for purifying extracellular vesicles have limitations such as induced aggregation, low yields, poor scalability, and one-dimensional separation process. Ion exchange chromatography, especially anion exchange chromatography, has been evaluated as a promising alternative for extracellular vesicle purification. However, cation exchange chromatography could also be applicable due to cationic domains on the vesicle surface. The choice of resin structure for cation exchange chromatography is critical, as the conventional-type cation exchanger only captures and elutes less than 20% of extracellular vesicles, while the tentacle-type resin can achieve comparable yields and superior purity when combined with conventional cation exchange resin.
JOURNAL OF CHROMATOGRAPHY A
(2023)
Article
Biochemistry & Molecular Biology
Jinxin Fan, Eduardo Barbieri, Shriarjun Shastry, Stefano Menegatti, Cristiana Boi, Ruben G. Carbonell
Summary: This study developed nonwoven ion-exchange membrane adsorbents for efficient purification of AAV2 from cell lysate and achieved high purity by utilizing a series of two different ion-exchange membranes.
Article
Biotechnology & Applied Microbiology
Saurabh Gautam, Dongyue Xin, Alan Pardo Garcia, Bart Spiesschaert
Summary: Purification of viruses, especially for therapeutic purposes, is a challenging task. This study presents a new single-step purification process for VSV-GP virus with high yields and purity. The purified virus has been characterized and demonstrated to have high purity and yield, making it suitable for various studies.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
