Using continuous chromatography methodology to achieve high-productivity and high-purity enrichment of charge variants for analytical characterization

Bigelow, Elizabeth; Song, Yuanli; Chen, Jie; Holstein, Melissa; Huang, Yunping; Duhamel, Lauren; Stone, Kelly D.; Furman, Ran; Li, Zheng Jian; Ghose, Sanchayita

doi:10.1016/j.chroma.2021.462008

Cited by 14 publications

(5 citation statements)

References 24 publications

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“…Researchers have previously proposed use of a multi-column setup for enrichment of charge variants. 24,30 Although complete displacement of acidic species may be desirable for some applications such as product characterization, it is not suitable for commercial processing. For biosimilar manufacturers, the primary objective is to match the charge variant profile to the innovator product.…”

Section: Resultsmentioning

confidence: 99%

A novel method for continuous chromatographic separation of monoclonal antibody charge variants by combining displacement mode chromatography and step elution

Anupa,

Bansode,

Kateja

et al. 2023

Biotechnology Progress

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Charge heterogeneity of monoclonal antibodies is considered a critical quality attribute and hence needs to be monitored and controlled by the manufacturer. Typically, this is accomplished via separation of charge variants on cation exchange chromatography (CEX) using a pH or conductivity based linear gradient elution. Although an effective approach, this is challenging particularly during continuous processing as creation of linear gradient during continuous processing adds to process complexity and can lead to deviations in product quality upon slightest changes in gradient formation. Moreover, the long length of elution gradient along with the required peak fractionation makes process integration difficult. In this study, we propose a novel approach for separation of charge variants during continuous CEX chromatography by utilizing a combination of displacement mode chromatography and salt‐based step elution. It has been demonstrated that while the displacement mode of chromatography enables control of acidic variants ≤26% in the CEX eluate, salt‐based step gradient elution manages basic charge variant ≤25% in the CEX eluate. The proposed approach has been successfully demonstrated using feed materials with varying compositions. On comparing the designed strategy with 2‐column concurrent (CC) chromatography, the resin specific productivity increased by 95% and resin utilization increased by 183% with recovery of main species >99%. Further, in order to showcase the amenability of the designed CEX method in continuous operation, the method was examined in our in‐house continuous mAb platform.

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Section: Resultsmentioning

confidence: 99%

A novel method for continuous chromatographic separation of monoclonal antibody charge variants by combining displacement mode chromatography and step elution

Anupa,

Bansode,

Kateja

et al. 2023

Biotechnology Progress

View full text Add to dashboard Cite

show abstract

“…Isolation of product-related impurities prior to characterization is currently performed utilizing inefficient and time-consuming procedures. The use of linear-scale RP or IEx chromatographic technique, either employing analytical LC or preparative systems coupled to fraction collectors, is a standard strategy [135,136]. These techniques rely on the linear range of the adsorption isotherm, achieving great resolution, minimizing components overlapping, and yielding high purity outcomes.…”

Section: N-rich Twin-column Continuous Chromatographymentioning

confidence: 99%

Sensitivity Enhancement for Separation-Based Analytical Techniques Utilizing Solid-Phase Enrichment Approaches and Analyte Derivatization for Trace Analysis in Various Matrices

et al. 2023

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Despite the fact that strong routine separation methodologies can give reliable specificity and validity at usual working pharmaceutical concentrations, they may fail at very low concentration levels. This poses considerable challenges for researchers investigating product purity and therapeutic drug monitoring. Sensitivity enhancement procedures are thus required to maximize the performance of separation techniques. Solid-phase extraction/solid-phase enrichment (SPE/SPEn) and pre-, post-, and in-column derivatization, as well as the use of sensitive detection devices, are the simplest strategies for improving sensitivity of separation-based analytical techniques. Large-volume injection of samples with online SPE/SPEn coupled with separation techniques increased sensitivity and improved detection as well as quantification limits without affecting peak shape and system performance. Although the primary purpose of derivatization is to improve sensitivity and selectivity, greener derivatization is growing in popularity and should be considered in analytical chemistry. In general, two strategies are essential for accomplishing greener derivatization goals. The first is the search for and use of ecologically acceptable derivatizing reagents, solvents, and reaction conditions. The second is miniaturization and automation of analytical methods. This review discusses significant advances in separation-based analytical techniques, specifically enrichment approaches and detector signal improvement for pharmaceutical quantification in various matrices at very low concentration levels. As a result of improved analytical systems setup in drug assays, the possibility of high-throughput analyses was also highlighted.

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“…The combination of continuous and countercurrent concepts coupled with the internal recycling and the possibility to use a solvent gradient program enables separation of ternary mixtures and allows to reach high product purities and elevated yields at the same time, outperforming the traditional single column processes, in many cases also in terms of productivity [ 125 , 126 ]. One of the first applications of the six-column MCSGP process concerned the purification of a 32 amino acids synthetic peptide, namely calcitonin, whose initial purity was 46%.…”

Section: Current Challenges and Future Perspectivesmentioning

confidence: 99%

Downstream Processing of Therapeutic Peptides by Means of Preparative Liquid Chromatography

et al. 2021

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The market of biomolecules with therapeutic scopes, including peptides, is continuously expanding. The interest towards this class of pharmaceuticals is stimulated by the broad range of bioactivities that peptides can trigger in the human body. The main production methods to obtain peptides are enzymatic hydrolysis, microbial fermentation, recombinant approach and, especially, chemical synthesis. None of these methods, however, produce exclusively the target product. Other species represent impurities that, for safety and pharmaceutical quality reasons, must be removed. The remarkable production volumes of peptide mixtures have generated a strong interest towards the purification procedures, particularly due to their relevant impact on the manufacturing costs. The purification method of choice is mainly preparative liquid chromatography, because of its flexibility, which allows one to choose case-by-case the experimental conditions that most suitably fit that particular purification problem. Different modes of chromatography that can cover almost every separation case are reviewed in this article. Additionally, an outlook to a very recent continuous chromatographic process (namely Multicolumn Countercurrent Solvent Gradient Purification, MCSGP) and future perspectives regarding purification strategies will be considered at the end of this review.

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Using continuous chromatography methodology to achieve high-productivity and high-purity enrichment of charge variants for analytical characterization

Cited by 14 publications

References 24 publications

A novel method for continuous chromatographic separation of monoclonal antibody charge variants by combining displacement mode chromatography and step elution

A novel method for continuous chromatographic separation of monoclonal antibody charge variants by combining displacement mode chromatography and step elution

Sensitivity Enhancement for Separation-Based Analytical Techniques Utilizing Solid-Phase Enrichment Approaches and Analyte Derivatization for Trace Analysis in Various Matrices

Downstream Processing of Therapeutic Peptides by Means of Preparative Liquid Chromatography

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