OpenLB
Open Library of Bioscience
Advanced Search
Methods in molecular biology (Clifton, N.J.)
2022;2531 15-47.

Capillary Electrophoresis-Mass Spectrometry (CE-MS) by Sheath-Flow Nanospray Interface and Its Use in Biopharmaceutical Applications.

Mei Han, Richard Smith, Dan A Rock
Author Information
  1. Mei Han: Pharmacokinetics and Drug Metabolism, Amgen Research, Amgen Inc., South San Francisco, CA, USA. hanm@amgen.com.
  2. Richard Smith: Pharmacokinetics and Drug Metabolism, Amgen Research, Amgen Inc., South San Francisco, CA, USA.
  3. Dan A Rock: Pharmacokinetics and Drug Metabolism, Amgen Research, Amgen Inc., South San Francisco, CA, USA.
PMID: 35941476 DOI: 10.1007/978-1-0716-2493-7_2

Abstract

Both capillary electrophoresis (CE) and mass spectrometry (MS) technologies are powerful analytical tools that have been used extensively in the characterization of biologics in the biopharmaceutical industry. The direct coupling of CE with MS is an attractive approach, in that the high separation capability of CE and the ultrasensitive detection and accurate identification performance of MS can be combined to provide a powerful system for the analysis of complex analytes. In this chapter, we discuss the detailed procedure of carrying out CE-MS analysis using a nano sheath-flow interface and its applications including intact mass analysis of monoclonal antibodies and fusion proteins, and a biotransformation study of two Fc-FGF21 molecules in a single-dose pharmacokinetic mice study. Optimization processes, including the finetuning of CE conditions and MS parameters, are illustrated in this chapter, with focuses on method robustness and assay reproducibility.

Keywords

Affinity capture Affinity purification Biotransformation CE-MS Capillary electrophoresis–mass spectrometry FGF21 Fusion protein Intact mass analysis Monoclonal antibody Time-of-flight

References

  1. Walsh G (2018) Biopharmaceutical benchmarks 2018. Nat Biotechnol 36:1136 [PMID: 30520869]
  2. Grilo AL, Mantalaris A (2019) The increasingly human and profitable monoclonal antibody market. Trends Biotechnol 37:9–16 [PMID: 29945725]
  3. Cruz E, Kayser V (2019) Monoclonal antibody therapy of solid tumors: clinical limitations and novel strategies to enhance treatment efficacy. Biologics 13:33–51 [PMID: 31118560]
  4. Awwad S, Angkawinitwong U (2018) Overview of antibody drug delivery. Pharmaceutics 10:83 [>PMCID: ]
  5. Audagnotto M, Dal Peraro M (2017) Protein post-translational modifications: in silico prediction tools and molecular modeling. Comput Struct Biotechnol J 15:307–319 [PMID: 28458782]
  6. Han M, Rock BM, Pearson JT, Rock DA (2016) Intact mass analysis of monoclonal antibodies by capillary electrophoresis-mass spectrometry. J Chromatogr B 1011:24–32 [DOI: 10.1016/j.jchromb.2015.12.045]
  7. Han M, Pearson JT, Wang Y, Winters D, Soto M, Rock DA, Rock BM (2017) Immunoaffinity capture coupled with capillary electrophoresis—mass spectrometry to study therapeutic protein stability in vivo. Anal Biochem 539:118–126 [PMID: 29029979]
  8. Creamer JS, Oborny NJ, Lunte SM (2014) Recent advances in the analysis of therapeutic proteins by capillary and microchip electrophoresis. Anal Methods 6:5427–5449 [PMID: 25126117]
  9. Zhao SS, Chen DD (2014) Applications of capillary electrophoresis in characterizing recombinant protein therapeutics. Electrophoresis 35:96–108 [PMID: 24123141]
  10. Sastre Torano J, Ramautar R, de Jong G (2019) Advances in capillary electrophoresis for the life sciences. J Chromatogr B 1118–1119:116–136 [DOI: 10.1016/j.jchromb.2019.04.020]
  11. Han M, Phan D, Nightlinger N, Taylor L, Jankhah S, Woodruff B, Yates Z, Freeman S, Guo A, Balland A, Pettit D (2006) Optimization of CE-SDS method for antibody separation based on multi-users experimental practices. Chromatographia 64:1–8 [DOI: 10.1365/s10337-006-0825-7]
  12. Salas-Solano O, Tomlinson B, Du S, Parker M, Strahan A, Ma S (2006) Optimization and validation of a quantitative capillary electrophoresis sodium dodecyl sulfate method for quality control and stability monitoring of monoclonal antibodies. Anal Chem 78:6583–6594 [PMID: 16970337]
  13. Han M, Guo A, Jochheim C, Zhang Y, Martinez T, Kodama P, Pettit D, Balland A (2007) Analysis of glycosylated type II Interleukin-1 receptor (IL-1R) by imaged capillary isoelectric focusing (i-cIEF). Chromatographia 66:969–976 [DOI: 10.1365/s10337-007-0338-z]
  14. Anderson CL, Wang Y, Rustandi RR (2012) Applications of imaged capillary isoelectric focussing technique in development of biopharmaceutical glycoprotein-based products. Electrophoresis 33:1538–1544 [PMID: 22736354]
  15. Gahoual R, Beck A, Leize-Wagner E, Francois YN (2016) Cutting-edge capillary electrophoresis characterization of monoclonal antibodies and related products. J Chromatogr B 1032:61–78 [DOI: 10.1016/j.jchromb.2016.05.028]
  16. Lechner A, Giorgetti J, Gahoual R, Beck A, Leize-Wagner E, Francois YN (2019) Insights from capillary electrophoresis approaches for characterization of monoclonal antibodies and antibody drug conjugates in the period 2016–2018. J Chromatogr B 1122–1123:1–17 [DOI: 10.1016/j.jchromb.2019.05.014]
  17. Kahle J, Watzig H (2018) Determination of protein charge variants with (imaged) capillary isoelectric focusing and capillary zone electrophoresis. Electrophoresis 39:2492–2511 [PMID: 29770965]
  18. Hamm M, Wang Y, Rustandi RR (2013) Characterization of N-linked glycosylation in a monoclonal antibody produced in NS0 cells using capillary electrophoresis with laser-induced fluorescence detection. Pharmaceuticals (Basel) 6:393–406 [DOI: 10.3390/ph6030393]
  19. Larsen MR, Trelle MB, Thingholm TE, Jensen ON (2006) Analysis of posttranslational modifications of proteins by tandem mass spectrometry. BioTechniques 40:790–798 [PMID: 16774123]
  20. Doll S, Burlingame AL (2015) Mass spectrometry-based detection and assignment of protein posttranslational modifications. ACS Chem Biol 10:63–71 [PMID: 25541750]
  21. Chait BT, Cadene M, Olinares PD, Rout MP, Shi Y (2016) Revealing higher order protein structure using mass spectrometry. J Am Soc Mass Spectrom 27:952–965 [PMID: 27080007]
  22. Liu CC, Zhang J, Dovichi NJ (2005) A sheath-flow nanospray interface for capillary electrophoresis/mass spectrometry. Rapid Commun Mass Spectrom 19:187–192 [PMID: 15593250]
  23. Gahoual R, Busnel J-M, Wolff P, François YN, Leize-Wagner E (2014) Novel sheathless CE-MS interface as an original and powerful infusion platform for nanoESI study: from intact proteins to high molecular mass noncovalent complexes. Anal Bioanal Chem 406:1029–1038 [PMID: 23881366]
  24. Moini M (2007) Simplifying CE−MS operation. 2. Interfacing low-flow separation techniques to mass spectrometry using a porous tip. Anal Chem 79:4241–4246 [PMID: 17447730]
  25. Sun L, Zhu G, Zhang Z, Mou S, Dovichi NJ (2015) Third-generation electrokinetically pumped sheath-flow nanospray interface with improved stability and sensitivity for automated capillary zone electrophoresis–mass spectrometry analysis of complex proteome digests. J Proteome Res 14:2312–2321 [PMID: 25786131]
  26. Smith RD, Barinaga CJ, Udseth HR (1988) Improved electrospray ionization interface for capillary zone electrophoresis-mass spectrometry. Anal Chem 60:1948–1952 [DOI: 10.1021/ac00169a022]
  27. Lindenburg PW, Haselberg R, Rozing G, Ramautar R (2015) Developments in interfacing designs for CE–MS: towards enabling tools for proteomics and metabolomics. Chromatographia 78:367–377 [DOI: 10.1007/s10337-014-2795-5]
  28. Zhu G, Sun L, Yan X, Dovichi NJ (2014) Stable, reproducible, and automated capillary zone electrophoresis-tandem mass spectrometry system with an electrokinetically pumped sheath-flow nanospray interface. Anal Chim Acta 810:94–98 [PMID: 24439510]
  29. Schiavone NM, Sarver SA, Sun L, Wojcik R, Dovichi NJ (2015) High speed capillary zone electrophoresis-mass spectrometry via an electrokinetically pumped sheath flow interface for rapid analysis of amino acids and a protein digest. J Chromatogr B 991:53–58 [DOI: 10.1016/j.jchromb.2015.04.001]
  30. Sun L, Hebert AS, Yan X, Zhao Y, Westphall MS, Rush MJ, Zhu G, Champion MM, Coon JJ, Dovichi NJ (2014) Over 10,000 peptide identifications from the HeLa proteome by using single-shot capillary zone electrophoresis combined with tandem mass spectrometry. Angew Chem Int Ed Engl 53:13931–13933 [PMID: 25346227]
  31. Stepanova S, Kasicka V (2016) Recent developments and applications of capillary and microchip electrophoresis in proteomic and peptidomic analyses. J Sep Sci 39:198–211 [PMID: 26497009]
  32. Kasicka V (2018) Recent developments in capillary and microchip electroseparations of peptides (2015-mid 2017). Electrophoresis 39:209–234 [PMID: 28836681]
  33. Dai J, Lamp J, Xia Q, Zhang Y (2018) Capillary isoelectric focusing-mass spectrometry method for the separation and online characterization of intact monoclonal antibody charge variants. Anal Chem 90:2246–2254 [PMID: 29272582]
  34. Zhu G, Sun L, Heidbrink-Thompson J, Kuntumalla S, Lin HY, Larkin CJ, McGivney JBT, Dovichi NJ (2016) Capillary zone electrophoresis tandem mass spectrometry detects low concentration host cell impurities in monoclonal antibodies. Electrophoresis 37:616–622 [PMID: 26530276]
  35. Zhao Y, Riley NM, Sun L, Hebert AS, Yan X, Westphall MS, Rush MJ, Zhu G, Champion MM, Mba Medie F, Champion PA, Coon JJ, Dovichi NJ (2015) Coupling capillary zone electrophoresis with electron transfer dissociation and activated ion electron transfer dissociation for top-down proteomics. Anal Chem 87:5422–5429 [PMID: 25893372]
  36. Han M, Rock BM, Pearson JT, Wang Y, Rock DA (2016) Therapeutic monoclonal antibody intact mass analysis by capillary electrophoresis–mass spectrometry. In: Xia JQ, Zhang L (eds) Capillary electrophoresis-mass spectrometry: therapeutic protein characterization. Springer International, Cham, pp 13–34 [DOI: 10.1007/978-3-319-46240-0_3]
  37. Zell M, Husser C, Staack RF, Jordan G, Richter WF, Schadt S, Pahler A (2016) In vivo biotransformation of the fusion protein Tetranectin-apolipoprotein A1 analyzed by ligand-binding mass spectrometry combined with quantitation by ELISA. Anal Chem 88:11670–11677 [PMID: 27934109]
  38. Kang L, Camacho RC, Li W, D'Aquino K, You S, Chuo V, Weng N, Jian W (2017) Simultaneous catabolite identification and quantitation of large therapeutic protein at the intact level by Immunoaffinity capture liquid chromatography-high-resolution mass spectrometry. Anal Chem 89:6065–6075 [PMID: 28457123]
  39. Kang L, Weng N, Jian W (2019) LC-MS bioanalysis of intact proteins and peptides. Biomed Chromatogr 34:e4633 [PMID: 31257628]
  40. Tibbitts J, Canter D, Graff R, Smith A, Khawli LA (2016) Key factors influencing ADME properties of therapeutic proteins: a need for ADME characterization in drug discovery and development. mAbs 8:229–245 [PMID: 26636901]
  41. Tumey LN, Rago B, Han X (2015) In vivo biotransformations of antibody-drug conjugates. Bioanalysis 7:1649–1664 [PMID: 26226313]
  42. Hager T, Spahr C, Xu J, Salimi-Moosavi H, Hall M (2013) Differential enzyme-linked immunosorbent assay and ligand-binding mass spectrometry for analysis of biotransformation of protein therapeutics: application to various FGF21 modalities. Anal Chem 85:2731–2738 [PMID: 23373459]
  43. Hecht R, Li Y-S, Sun J, Belouski E, Hall M, Hager T, Yie J, Wang W, Winters D, Smith S, Spahr C, Tam L-T, Shen Z, Stanislaus S, Chinookoswong N, Lau Y, Sickmier A, Michaels ML, Boone T, Véniant MM, Xu J (2012) Rationale-based engineering of a potent long-acting FGF21 analog for the treatment of type 2 diabetes. PLoS One 7:e49345 [PMID: 23209571]

MeSH Term

Animals
Antibodies, Monoclonal
Biological Products
Electrophoresis, Capillary
Mass Spectrometry
Mice
Reproducibility of Results
Spectrometry, Mass, Electrospray Ionization

Chemicals

Antibodies, Monoclonal
Biological Products
Journal Article
Article has an altmetric score of 7

Word Cloud

Created with Highcharts 10.0.0CEMSanalysismassCE-MSspectrometrypowerfulchapterincludingstudyCapillaryAffinitycapillaryelectrophoresistechnologiesanalyticaltoolsusedextensivelycharacterizationbiologicsbiopharmaceuticalindustrydirectcouplingattractiveapproachhighseparationcapabilityultrasensitivedetectionaccurateidentificationperformancecancombinedprovidesystemcomplexanalytesdiscussdetailedprocedurecarryingusingnanosheath-flowinterfaceapplicationsintactmonoclonalantibodiesfusionproteinsbiotransformationtwoFc-FGF21moleculessingle-dosepharmacokineticmiceOptimizationprocessesfinetuningconditionsparametersillustratedfocusesmethodrobustnessassayreproducibilityElectrophoresis-MassSpectrometrySheath-FlowNanosprayInterfaceUseBiopharmaceuticalApplicationscapturepurificationBiotransformationelectrophoresis–massFGF21FusionproteinIntactMonoclonalantibodyTime-of-flight

Similar Articles

Cited By

No available data.