Automated ion exchange chromatography screening combined with in silico multifactorial simulation for efficient method development and purification of biopharmaceutical targets.
Biopharmaceutical
Computer-assisted modeling
Fraction collection
Ion exchange chromatography
Multidimensional liquid chromatography
Screening
Journal
Analytical and bioanalytical chemistry
ISSN: 1618-2650
Titre abrégé: Anal Bioanal Chem
Pays: Germany
ID NLM: 101134327
Informations de publication
Date de publication:
May 2022
May 2022
Historique:
received:
22
12
2021
accepted:
15
02
2022
revised:
10
02
2022
pubmed:
21
4
2022
medline:
27
4
2022
entrez:
20
4
2022
Statut:
ppublish
Résumé
Bioprocess development of increasingly challenging therapeutics and vaccines requires a commensurate level of analytical innovation to deliver critical assays across functional areas. Chromatography hyphenated to numerous choices of detection has undeniably been the preferred analytical tool in the pharmaceutical industry for decades to analyze and isolate targets (e.g., APIs, intermediates, and byproducts) from multicomponent mixtures. Among many techniques, ion exchange chromatography (IEX) is widely used for the analysis and purification of biopharmaceuticals due to its unique selectivity that delivers distinctive chromatographic profiles compared to other separation modes (e.g., RPLC, HILIC, and SFC) without denaturing protein targets upon isolation process. However, IEX method development is still considered one of the most challenging and laborious approaches due to the many variables involved such as elution mechanism (via salt, pH, or salt-mediated-pH gradients), stationary phase's properties (positively or negatively charged; strong or weak ion exchanger), buffer type and ionic strength as well as pH choices. Herein, we introduce a new framework consisting of a multicolumn IEX screening in conjunction with computer-assisted simulation for efficient method development and purification of biopharmaceuticals. The screening component integrates a total of 12 different columns and 24 mobile phases that are sequentially operated in a straightforward automated fashion for both cation and anion exchange modes (CEX and AEX, respectively). Optimal and robust operating conditions are achieved via computer-assisted simulation using readily available software (ACD Laboratories/LC Simulator), showcasing differences between experimental and simulated retention times of less than 0.5%. In addition, automated fraction collection is also incorporated into this framework, illustrating the practicality and ease of use in the context of separation, analysis, and purification of nucleotides, peptides, and proteins. Finally, we provide examples of the use of this IEX screening as a framework to identify efficient first dimension (
Identifiants
pubmed: 35441858
doi: 10.1007/s00216-022-03982-z
pii: 10.1007/s00216-022-03982-z
doi:
Substances chimiques
Biological Products
0
Peptides
0
Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
3581-3591Informations de copyright
© 2022. Merck & Co., Inc., Kenilworth, NJ, USA and its affiliates and Davy Guillarme under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
Références
Regalado EL, Haidar Ahmad IA, Bennett R, D’Atri V, Makarov AA, Humphrey GR, et al. The emergence of universal chromatographic methods in the research and development of new drug substances. Acc Chem Res. 2019;52(7):1990–2002.
pubmed: 31198042
doi: 10.1021/acs.accounts.9b00068
Johnson DE. Biotherapeutics: challenges and opportunities for predictive toxicology of monoclonal antibodies. Int J Mol Sci. 2018;19(11):3685.
pmcid: 6274697
doi: 10.3390/ijms19113685
Gautam A, Pan X. The changing model of big pharma: impact of key trends. Drug Discovery Today. 2016;21(3):379–84.
pubmed: 26477304
doi: 10.1016/j.drudis.2015.10.002
Fekete S, Guillarme D, Sandra P, Sandra K. Chromatographic, electrophoretic, and mass spectrometric methods for the analytical characterization of protein biopharmaceuticals. Anal Chem. 2016;88(1):480–507.
pubmed: 26629607
doi: 10.1021/acs.analchem.5b04561
Haidar Ahmad IA, Bennett R, Makey D, Shchurik V, Lhotka H, Mann BF, et al. In silico method development for the reversed-phase liquid chromatography separation of proteins using chaotropic mobile phase modifiers. J Chromatogr B. 2021;1173:122587.
doi: 10.1016/j.jchromb.2021.122587
Camperi J, Goyon A, Guillarme D, Zhang K, Stella C. Multi-dimensional LC-MS: the next generation characterization of antibody-based therapeutics by unified online bottom-up, middle-up and intact approaches. Analyst. 2021;146(3):747–69.
pubmed: 33410843
doi: 10.1039/D0AN01963A
Goyon A, D’Atri V, Bobaly B, Wagner-Rousset E, Beck A, Fekete S, et al. Protocols for the analytical characterization of therapeutic monoclonal antibodies. I – non-denaturing chromatographic techniques. J Chromatogr B. 2017;1058:73–84.
doi: 10.1016/j.jchromb.2017.05.010
Losacco GL, DaSilva JO, Liu J, Regalado EL, Veuthey J-L, Guillarme D. Expanding the range of sub/supercritical fluid chromatography: advantageous use of methanesulfonic acid in water-rich modifiers for peptide analysis. J Chromatogr A. 2021;1642:462048.
pubmed: 33744606
doi: 10.1016/j.chroma.2021.462048
Goyon A, Zhang K. Characterization of antisense oligonucleotide impurities by ion-pairing reversed-phase and anion exchange chromatography coupled to hydrophilic interaction liquid chromatography/mass spectrometry using a versatile two-dimensional liquid chromatography setup. Anal Chem. 2020;92(8):5944–51.
pubmed: 32191031
doi: 10.1021/acs.analchem.0c00114
Bennett R, Biba M, Liu J, Haidar Ahmad IA, Hicks MB, Regalado EL. Enhanced fluidity liquid chromatography: a guide to scaling up from analytical to preparative separations. J Chromatogr A. 2019;1595:190–8.
pubmed: 30803788
doi: 10.1016/j.chroma.2019.02.017
Ikegami T. Hydrophilic interaction chromatography for the analysis of biopharmaceutical drugs and therapeutic peptides: a review based on the separation characteristics of the hydrophilic interaction chromatography phases. J Sep Sci. 2019;42(1):130–213.
pubmed: 30461188
doi: 10.1002/jssc.201801074
Jaag S, Shirokikh M, Lämmerhofer M. Charge variant analysis of protein-based biopharmaceuticals using two-dimensional liquid chromatography hyphenated to mass spectrometry. J Chromatogr A. 2021;1636:461786.
pubmed: 33326927
doi: 10.1016/j.chroma.2020.461786
Periat A, Fekete S, Cusumano A, Veuthey J-L, Beck A, Lauber M, et al. Potential of hydrophilic interaction chromatography for the analytical characterization of protein biopharmaceuticals. J Chromatogr A. 2016;1448:81–92.
pubmed: 27131959
doi: 10.1016/j.chroma.2016.04.056
Piestansky J, Barath P, Majerova P, Galba J, Mikus P, Kovacech B, et al. A simple and rapid LC-MS/MS and CE-MS/MS analytical strategy for the determination of therapeutic peptides in modern immunotherapeutics and biopharmaceutics. J Pharm Biomed Anal. 2020;189:113449.
pubmed: 32622303
doi: 10.1016/j.jpba.2020.113449
Fekete S, Beck A, Veuthey J-L, Guillarme D. Ion-exchange chromatography for the characterization of biopharmaceuticals. J Pharm Biomed Anal. 2015;113:43–55.
pubmed: 25800161
doi: 10.1016/j.jpba.2015.02.037
Abou El Azm N, Fleita D, Rifaat D, Mpingirika EZ, Amleh A, El-Sayed MMH. Production of bioactive compounds from the sulfated polysaccharides extracts of Ulva lactuca: post-extraction enzymatic hydrolysis followed by ion-exchange chromatographic fractionation. Molecules. 2019;24(11):2132.
pmcid: 6600532
doi: 10.3390/molecules24112132
Leblanc Y, Bihoreau N, Chevreux G. Characterization of human serum albumin isoforms by ion exchange chromatography coupled on-line to native mass spectrometry. J Chromatogr B. 2018;1095:87–93.
doi: 10.1016/j.jchromb.2018.07.014
McGinnis AC, Cummings BS, Bartlett MG. Ion exchange liquid chromatography method for the direct determination of small ribonucleic acids. Anal Chim Acta. 2013;799:57–67.
pubmed: 24091375
doi: 10.1016/j.aca.2013.08.040
Mommen GPM, Meiring HD, Heck AJR, de Jong APJM. Mixed-bed ion exchange chromatography employing a salt-free pH gradient for improved sensitivity and compatibility in MudPIT. Anal Chem. 2013;85(14):6608–16.
pubmed: 23772827
doi: 10.1021/ac400995e
Bertoletti L, Regazzoni L, Aldini G, Colombo R, Abballe F, Caccialanza G, et al. Separation and characterisation of beta2-microglobulin folding conformers by ion-exchange liquid chromatography and ion-exchange liquid chromatography–mass spectrometry. Anal Chim Acta. 2013;771:108–14.
pubmed: 23522119
doi: 10.1016/j.aca.2013.01.058
Haidar Ahmad IA, Shchurik V, Nowak T, Mann BF, Regalado EL. Introducing multifactorial peak crossover in analytical and preparative chromatography via computer-assisted modeling. Anal Chem. 2020;92(19):13443–51.
pubmed: 32786491
doi: 10.1021/acs.analchem.0c02807
Ahmed S, Atia NN, Rageh AH. Selectivity enhanced cation exchange chromatography for simultaneous determination of peptide variants. Talanta. 2019;199:347–54.
pubmed: 30952269
doi: 10.1016/j.talanta.2019.02.082
Verscheure L, Cerdobbel A, Sandra P, Lynen F, Sandra K. Monoclonal antibody charge variant characterization by fully automated four-dimensional liquid chromatography-mass spectrometry. J Chromatogr A. 2021;1653:462409.
pubmed: 34325295
doi: 10.1016/j.chroma.2021.462409
Gstöttner C, Klemm D, Haberger M, Bathke A, Wegele H, Bell C, et al. Fast and automated characterization of antibody variants with 4D HPLC/MS. Anal Chem. 2018;90(3):2119–25.
pubmed: 29264912
doi: 10.1021/acs.analchem.7b04372
Füssl F, Trappe A, Carillo S, Jakes C, Bones J. Comparative elucidation of cetuximab heterogeneity on the intact protein level by cation exchange chromatography and capillary electrophoresis coupled to mass spectrometry. Anal Chem. 2020;92(7):5431–8.
pubmed: 32105056
doi: 10.1021/acs.analchem.0c00185
Schiavone NM, Bennett R, Hicks MB, Pirrone GF, Regalado EL, Mangion I, et al. Evaluation of global conformational changes in peptides and proteins following purification by supercritical fluid chromatography. J Chromatogr B. 2019;1110–1111:94–100.
doi: 10.1016/j.jchromb.2019.02.012
Li Z, Wang Q, Wang Y, Wang K, Liu Z, Zhang W, et al. An efficient approach based on basic strong cation exchange chromatography for enriching methylated peptides with high specificity for methylproteomics analysis. Anal Chim Acta. 2021;1161:338467.
pubmed: 33896563
doi: 10.1016/j.aca.2021.338467
Patel BA, Pinto NDS, Gospodarek A, Kilgore B, Goswami K, Napoli WN, et al. On-line ion exchange liquid chromatography as a process analytical technology for monoclonal antibody characterization in continuous bioprocessing. Anal Chem. 2017;89(21):11357–65.
pubmed: 28981255
doi: 10.1021/acs.analchem.7b02228
Tsay F-R, Haidar Ahmad IA, Henderson D, Schiavone N, Liu Z, Makarov AA, et al. Generic anion-exchange chromatography method for analytical and preparative separation of nucleotides in the development and manufacture of drug substances. J Chromatogr A. 2019;1587:129–35.
pubmed: 30591245
doi: 10.1016/j.chroma.2018.12.018
Yan Y, Liu AP, Wang S, Daly TJ, Li N. Ultrasensitive characterization of charge heterogeneity of therapeutic monoclonal antibodies using strong cation exchange chromatography coupled to native mass spectrometry. Anal Chem. 2018;90(21):13013–20.
pubmed: 30280893
doi: 10.1021/acs.analchem.8b03773
Losacco GL, Wang H, Ahmad IH, DaSilva J, Makarov AA, Mangion I, et al. Enantioselective UHPLC screening combined with in silico modeling for streamlined development of ultrafast enantiopurity assays. Anal Chem. 2022;94(3):1804–12.
pubmed: 34931812
doi: 10.1021/acs.analchem.1c04585
Haidar Ahmad IA, Makey DM, Wang H, Shchurik V, Singh AN, Stoll DR, et al. In silico multifactorial modeling for streamlined development and optimization of two-dimensional liquid chromatography. Anal Chem. 2021;93(33):11532–9.
pubmed: 34375071
doi: 10.1021/acs.analchem.1c01970
Wang H, Herderschee HR, Bennett R, Potapenko M, Pickens CJ, Mann BF, et al. Introducing online multicolumn two-dimensional liquid chromatography screening for facile selection of stationary and mobile phase conditions in both dimensions. J Chromatogr A. 2020;1622:460895.
pubmed: 32408991
doi: 10.1016/j.chroma.2020.460895
Barhate CL, Joyce LA, Makarov AA, Zawatzky K, Bernardoni F, Schafer WA, et al. Ultrafast chiral separations for high throughput enantiopurity analysis. Chem Commun. 2017;53(3):509–12.
doi: 10.1039/C6CC08512A
Haidar Ahmad IA, Chen W, Halsey HM, Klapars A, Limanto J, Pirrone GF, et al. Multi-column ultra-high performance liquid chromatography screening with chaotropic agents and computer-assisted separation modeling enables process development of new drug substances. Analyst. 2019;144(9):2872–80.
pubmed: 30830135
doi: 10.1039/C8AN02499E
D’Atri V, Murisier A, Fekete S, Veuthey J-L, Guillarme D. Current and future trends in reversed-phase liquid chromatography-mass spectrometry of therapeutic proteins. TrAC Trends Anal Chem. 2020;130:115962.
doi: 10.1016/j.trac.2020.115962
De Pra M, Greco G, Krajewski MP, Martin MM, George E, Bartsch N, et al. Effects of titanium contamination caused by iron-free high-performance liquid chromatography systems on peak shape and retention of drugs with chelating properties. J Chromatogr A. 2020;1611:460619.
pubmed: 31668415
doi: 10.1016/j.chroma.2019.460619
Bennett R, Cohen RD, Wang H, Pereira T, Haverick MA, Loughney JW, et al. A selective plate-based assay for trace EDTA analysis via boron trifluoride-methanol derivatization UHPLC-QqQ-MS/MS enabling biologic and vaccine processes. Anal Chem. 2022;94(3):1678–85.
pubmed: 34928586
doi: 10.1021/acs.analchem.1c04224
Guichard N, Fekete S, Guillarme D, Bonnabry P, Fleury-Souverain S. Computer-assisted UHPLC-MS method development and optimization for the determination of 24 antineoplastic drugs used in hospital pharmacy. J Pharm Biomed Anal. 2019;164:395–401.
pubmed: 30439666
doi: 10.1016/j.jpba.2018.11.014
Haidar Ahmad IA, Kiffer A, Barrientos RC, Losacco GL, Singh A, Shchurik V, Wang H, Mangion I, Regalado EL, et al. In Silico Method Development of Achiral and Chiral Tandem Column Reversed-phase Liquid Chromatography for Multicomponent Pharmaceutical Mixtures. Anal Chem. 2022;94:4065–71.
pubmed: 35199987
doi: 10.1021/acs.analchem.1c05551
Makey DM, Shchurik V, Wang H, Lhotka HR, Stoll DR, Vazhentsev A, et al. Mapping the separation landscape in two-dimensional liquid chromatography: blueprints for efficient analysis and purification of pharmaceuticals enabled by computer-assisted modeling. Anal Chem. 2021;93(2):964–72.
pubmed: 33301312
doi: 10.1021/acs.analchem.0c03680
Bennett R, Haidar Ahmad IA, DaSilva J, Figus M, Hullen K, Tsay F-R, et al. Mapping the separation landscape of pharmaceuticals: rapid and efficient scale-up of preparative purifications enabled by computer-assisted chromatographic method development. Org Process Res Dev. 2019;23(12):2678–84.
doi: 10.1021/acs.oprd.9b00351