Keeping up with a Quickly Diversifying Pharmaceutical Landscape DOI Creative Commons
Alexandre Goyon

ACS Measurement Science Au, Journal Year: 2024, Volume and Issue: 4(6), P. 615 - 619

Published: Sept. 24, 2024

Small molecules and antibodies have dominated the pharmaceutical landscape for decades. However, limitations associated with therapeutic targets deemed "undruggable" progress in biology chemistry led to blossoming of drug modalities approaches. In 2023, a high number 9 oligonucleotide peptide products were approved by Food Drug Administration (FDA), accounting 16% all drugs approved. Additionally, first time, clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 gene therapy product was treatment sickle cell disease. New possess wide range physicochemical properties structures, which complicates their analytical characterization. Impurities are formed at each step solid phase synthesis during shelf life. Longer chain lengths lead higher closely related impurities that become increasingly more difficult separate from full-length product. Chemical modifications such as phosphorothioates (PS) result presence diastereomers, often require orthogonal methods profiling strategies prevent interference separation achiral impurities. In-vitro produced mRNA plasmid DNA also present variety quality attributes need be determined, polyA tail length or capping efficiency. Analytical challenges arise modality physiochemical attributes, fast turnaround times, heightened level characterization needed enable data-driven decisions early development process. This perspective provides author's views on lessons learned employed recent years.

Language: Английский

The Role of mD-LC–MS for Antibody Analysis in the Biopharmaceutical Industry DOI
Katrin Heinrich,

Saban Özipek,

Sina Hoelterhoff

et al.

Deleted Journal, Journal Year: 2025, Volume and Issue: unknown, P. 4 - 10

Published: Jan. 3, 2025

The field of therapeutic antibodies has witnessed remarkable growth in recent years, with novel biotherapeutics formats continually emerging. As the complexity these increases, so does demand for robust analytical methods to characterize them. In this article, we explore transformative impact multidimensional liquid chromatography–mass spectrometry (mD-LC–MS) technology on antibody characterization, a focus specific performance enhancements. This article explores challenges faced by traditional chromatographic and discusses how mD-LC–MS is changing way evaluate high chromatography (HPLC) peaks assess performance. Specifically, highlights enables precise peak identification, rapid characterization complex antibodies, early detection post-translational modifications improve assessment potential critical quality attributes (pCQAs) optimize overall workflow. advance led significant time savings reduction artificial induced consumption, thereby reducing risk sample during analysis.

Language: Английский

Citations

1
Evaluation of ion pairing reversed-phase liquid chromatography for the separation of large RNA molecules DOI Creative Commons
Jonathan Maurer,

Camille Malburet,

Marc François-Heude

et al.

Journal of Chromatography A, Journal Year: 2024, Volume and Issue: 1740, P. 465574 - 465574

Published: Dec. 2, 2024

The rapid development of mRNA-based therapeutics, especially post-COVID-19, has necessitated the precise characterization mRNA quality attributes, including sequence integrity. Ion-pairing reversed-phase liquid chromatography (IP-RPLC) been widely accepted as a reference method for small oligonucleotides. Some studies have already investigated use IP-RPLC RNA, but no systematic approach developed to assess impact ion-pairing agents (IPAs) on separation large RNA molecules. This study addresses this gap by investigating potential and molecules, with specific focus optimizing IPAs enhance retention selectivity. Thirteen different IPAs, varying in hydrophobicity, were systematically tested using supermacroporous polymeric (divinylbenzene) column very broad pore size range under various conditions, temperatures, pH, IPA concentrations. results demonstrate that moderately hydrophobic provide superior resolution species up 6000 nucleotides. An optimized combination 100 mM butylammonium acetate 50 tripropylammonium achieved best overall separation, significantly improving 35% compared individual IPAs. also identifies optimal conditions mobile phase pH 7.0, acetonitrile organic solvent, temperature 65 °C. In second step, solution increase nucleotides thereby separate nucleic acids ranging from 1 allowing characterize IVT-mRNA differing length their integrity fragmentation or monitor presence in-process impurities (nucleotides) was combining two LC columns. These findings analytical toolbox evaluating critical attributes supporting reliable efficient RNA-based therapeutics.

Language: Английский

Citations

4
Ion exchange chromatography of biotherapeutics: Fundamental principles and advanced approaches DOI Creative Commons
Mateusz Imiołek, Szabolcs Fekete, Serge Rudaz

et al.

Journal of Chromatography A, Journal Year: 2025, Volume and Issue: 1742, P. 465672 - 465672

Published: Jan. 9, 2025

Ion exchange chromatography (IEX) is an important analytical technique for the characterization of biotechnology-derived products, such as monoclonal antibodies (mAbs) and more recently, cell gene therapy products messenger ribonucleic acid (mRNA) adeno-associated viruses (AAVs). This review paper first outlines basic principles separation mechanisms IEX charge variant biotherapeutics, examines different elution modes based on salt or pH gradients. It then highlights several recent trends when applying including: i) effective use gradients, ii) improvement selectivity by using organic solvents in mobile phase, multi-step combining ion pairing exchange, iii) increase throughput ultra-short columns automated screening conditions. The also discusses incorporation into multidimensional liquid setups, integrating it with other chromatographic dimensions analysis complex biotherapeutic products. covers coupling mass spectrometry (MS), mobility (IMS), multi-angle light scattering (MALS) to identify various species contained samples. In conclusion, considered today essential toolbox quality control offers a unique mechanism can be coupled highly informative detectors, MS MALS.

Language: Английский

Citations

0
Advanced two-dimensional liquid chromatography workflow for enhanced resolution of protein components in Indian Cobra (Naja naja) venom using hydrophobic interaction -reverse phase chromatography coupled with mass spectrometry DOI Creative Commons
Sunil Kumar, Kunal Krishna, Anurag S. Rathore

et al.

Journal of Chromatography Open, Journal Year: 2025, Volume and Issue: unknown, P. 100211 - 100211

Published: March 1, 2025

Language: Английский

Citations

0
Establishing Multi-Dimensional LC-MS Systems for Versatile Workflows to Analyze Therapeutic Antibodies at Different Molecular Levels in Routine Operations DOI Creative Commons
Katrin Heinrich, Sina Hoelterhoff, Saban Oezipek

et al.

Pharmaceuticals, Journal Year: 2025, Volume and Issue: 18(3), P. 401 - 401

Published: March 12, 2025

Background/Objectives: Multi-dimensional liquid chromatography coupled with mass spectrometry (mD-LC-MS) has emerged as a powerful technique for the in-depth characterization of biopharmaceuticals by assessing chromatographically resolved product variants in streamlined and semi-automated manner. The study aims to demystify enhance accessibility this but inherently complex detailing robust user-friendly instrument platform, allowing analysts switch seamlessly between intact, subunit, peptide mapping workflows. Methods: Starting from commercially available Two-Dimensional Liquid Chromatography (2D-LC) system, we introduce specific hardware software extensions leading two versatile mD-LC-MS setups, slightly different configurations. technique’s efficacy is demonstrated through case on cation exchange method charge bispecific antibody, isolating peak(s) interest, followed online sample processing, including reduction enzymatic digestion, subsequently analysis. Results: accuracy reproducibility both setups proposed were successfully tested. Despite peak patterns first dimension, systems equally effective identifying quantifying underlying species. This highlights routine usability technology (ultra) high-performance (UHPLC) therapeutic biomolecule. Conclusions: reliability underscore practicality use biopharmaceutical Our detailed description insights simplify access advanced broader scientific community, regardless expertise level, lower entry barrier its various research industrial settings.

Language: Английский

Citations

0
Selected new approaches and future perspectives in liquid chromatography for the analysis of emerging modalities DOI Creative Commons
Szabolcs Fekete, Mateusz Imiołek, Fabrice Gritti

et al.

European Journal of Pharmaceutical Sciences, Journal Year: 2025, Volume and Issue: unknown, P. 107101 - 107101

Published: April 1, 2025

Emerging biopharmaceutical modalities, such as genetic medicines and RNA therapies, offer transformative potential for treating previously intractable diseases. However, these complex drugs present unique analytical challenges due to their intricate structures, sophisticated manufacturing processes, modality-specific product quality attributes. Liquid chromatography (LC) has emerged a versatile tool addressing challenges, enabling precise characterization control strategies. This review highlights recent advancements in LC technologies, including low-adsorption hardware, ultra-wide pore size exclusion (SEC) columns, innovative separation modes slalom pressure-enhanced liquid (PELC). These developments tackle issues non-specific adsorption, carryover, inadequate selectivity while improving resolution robustness large biomolecules like mRNA, adeno-associated viruses (AAVs), lipid nanoparticles (LNPs). Novel approaches, tandem SEC systems, gradient dual stationary phase gradients, further expand the scope of techniques by enhancing separations diverse analyte sizes complexities. Additionally, practical innovations bracketed injection methods new enzymatic tools oligo-mapping improve reproducibility, efficiency, confidence sequence analysis. not only address current limitations but also pave way regulatory-compliant which will support broader adoption both discovery settings. As field continues evolve, are poised play pivotal role ensuring safety, efficacy, consistency next-generation therapeutics.

Language: Английский

Citations

0
2D-CEX–FcRn–MS to Study Structure/Function Relation of mAb Charge Variants DOI
Liesa Verscheure,

Isabel Vandenheede,

Eline De Rore

et al.

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(45), P. 18122 - 18131

Published: Oct. 29, 2024

The automated elucidation of the interplay between monoclonal antibody (mAb) structure and function using two-dimensional liquid chromatography-mass spectrometry (2D-LC-MS) is reported. Charge variants, induced through forced degradation, are resolved by first-dimension (

Language: Английский

Citations

1
Keeping up with a Quickly Diversifying Pharmaceutical Landscape DOI Creative Commons
Alexandre Goyon

ACS Measurement Science Au, Journal Year: 2024, Volume and Issue: 4(6), P. 615 - 619

Published: Sept. 24, 2024

Small molecules and antibodies have dominated the pharmaceutical landscape for decades. However, limitations associated with therapeutic targets deemed "undruggable" progress in biology chemistry led to blossoming of drug modalities approaches. In 2023, a high number 9 oligonucleotide peptide products were approved by Food Drug Administration (FDA), accounting 16% all drugs approved. Additionally, first time, clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 gene therapy product was treatment sickle cell disease. New possess wide range physicochemical properties structures, which complicates their analytical characterization. Impurities are formed at each step solid phase synthesis during shelf life. Longer chain lengths lead higher closely related impurities that become increasingly more difficult separate from full-length product. Chemical modifications such as phosphorothioates (PS) result presence diastereomers, often require orthogonal methods profiling strategies prevent interference separation achiral impurities. In-vitro produced mRNA plasmid DNA also present variety quality attributes need be determined, polyA tail length or capping efficiency. Analytical challenges arise modality physiochemical attributes, fast turnaround times, heightened level characterization needed enable data-driven decisions early development process. This perspective provides author's views on lessons learned employed recent years.

Language: Английский

Citations

0