CoChat
Sign Up
Cited by 2

The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.

Micro simulated moving bed chromatography-mass spectrometry as a continuous on-line process analytical tool

Diehm, Juliane, Witting, Lennart, Kirschhöfer, Frank, Brenner-Weiß, Gerald, Franzreb, Matthias·2024·Analytical and Bioanalytical Chemistry
DOI 10.1007/s00216-023-05023-9PMC PMC10761468
Open this paper in CoChat — free account

Abstract

Continuous manufacturing is becoming increasingly important in the (bio-)pharmaceutical industry, as more product can be produced in less time and at lower costs. In this context, there is a need for powerful continuous analytical tools. Many established off-line analytical methods, such as mass spectrometry (MS), are hardly considered for process analytical technology (PAT) applications in biopharmaceutical processes, as they are limited to at-line analysis due to the required sample preparation and the associated complexity, although they would provide a suitable technique for the assessment of a wide range of quality attributes. In this study, we investigated the applicability of a recently developed micro simulated moving bed chromatography system (µSMB) for continuous on-line sample preparation for MS. As a test case, we demonstrate the continuous on-line MS measurement of a protein solution (myoglobin) containing Tris buffer, which interferes with ESI-MS measurements, by continuously exchanging this buffer with a volatile ammonium acetate buffer suitable for MS measurements. The integration of the µSMB significantly increases MS sensitivity by removing over 98% of the buffer substances. Thus, this study demonstrates the feasibility of on-line µSMB-MS, providing a versatile PAT tool by combining the detection power of MS for various product attributes with all the advantages of continuous on-line analytics. Graphical abstract

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1007/s00216-023-05023-9

Cites

1 of 1 references verified0 citation errors · 0 not checkable

Statistics

Figures

Fig. 1 Instrumentation and piping diagram of the used µSMB setup. The conductivity sensor in the raffinate stream is only used when the µSMB is not coupled to the MS
Fig. 1 Instrumentation and piping diagram of the used µSMB setup. The conductivity sensor in the raffinate stream is only used when the µSMB is not coupled to the MSCC BY · PubMed Central
Fig. 2 Measurement of Mb in native and non-native state. (a) background spectrum of 10 mM NH4OAc buffer and spectra of Mb under native and non-native measurement conditions. (b) Protein reconstruction of the Mb spectra from (a). (c) Close up of the Mb main peak with isotopic peaks and sodium adducts. In (a) and (c), charge states are assigned in parenthesis after m/z labels. In (a) and (b), peaks of holoMb are marked with white squares, of apoMb with black circles, and of heme with white circles
Fig. 2 Measurement of Mb in native and non-native state. (a) background spectrum of 10 mM NH4OAc buffer and spectra of Mb under native and non-native measurement conditions. (b) Protein reconstruction of the Mb spectra from (a). (c) Close up of the Mb main peak with isotopic peaks and sodium adducts. In (a) and (c), charge states are assigned in parenthesis after m/z labels. In (a) and (b), peaks of holoMb are marked with white squares, of apoMb with black circles, and of heme with white circlesCC BY · PubMed Central
Fig. 3 Influence of Tris buffer on MS detection of Mb. (a) Mass spectra of solutions containing 10 µg/mL Mb with increasing Tris concentrations. Charge states are assigned in parenthesis after m/z labels. (b) Normalized Tris signal (m/z 593.23) and amount of detected holoMb in dependence of Tris concentration. (c) Influence of Tris concentration on detected percentage of apoMb and heme group. The detected amount of apo/holoMb was calculated by integration of the respective peaks in the protein reconstruction chromatogram
Fig. 3 Influence of Tris buffer on MS detection of Mb. (a) Mass spectra of solutions containing 10 µg/mL Mb with increasing Tris concentrations. Charge states are assigned in parenthesis after m/z labels. (b) Normalized Tris signal (m/z 593.23) and amount of detected holoMb in dependence of Tris concentration. (c) Influence of Tris concentration on detected percentage of apoMb and heme group. The detected amount of apo/holoMb was calculated by integration of the respective peaks in the protein reconstruction chromatogramCC BY · PubMed Central
Fig. 4 Tris concentrations in the raffinate and extract streams during a µSMB experiment with 100 mM Tris in the feed solution
Fig. 4 Tris concentrations in the raffinate and extract streams during a µSMB experiment with 100 mM Tris in the feed solutionCC BY · PubMed Central
Fig. 5 Comparison of the different in- and outlet flow rates without (a) and with (b) direct coupling of the raffinate stream to the ESI interface of the MS system
Fig. 5 Comparison of the different in- and outlet flow rates without (a) and with (b) direct coupling of the raffinate stream to the ESI interface of the MS systemCC BY · PubMed Central
Fig. 6 Normalized signals of Mb, heme group, and Tris (a) during the entire first µSMB-MS experiment (run 1); (b) for the third process cycle at every Tris concentration (marked with black boxes in (a))
Fig. 6 Normalized signals of Mb, heme group, and Tris (a) during the entire first µSMB-MS experiment (run 1); (b) for the third process cycle at every Tris concentration (marked with black boxes in (a))CC BY · PubMed Central
Fig. 7 (a) Normalized Tris signal as a function of the Tris concentration in the feed of the µSMB experiments. Depicted values are the mean values of the third cycle of each concentration. The Tris concentrations of the calibrations were multiplied with the expected desalting level of 98% in order to match the x-axis. (b) Detected apoMb percentage and detected amount of Mb as a function of the feed Tris concentration during the experiments. The values are mean values of the third cycle of the µSMB process for each concentration. (c) Detected apoMb percentage and Tris signal level for three consecutive cycles (4–6) of µSMB run 2 with a constant Tris concentration of 10 mM in the feed stream. The apoMb level was calculated for each switching interval, as indicated by the dotted vertical lines
Fig. 7 (a) Normalized Tris signal as a function of the Tris concentration in the feed of the µSMB experiments. Depicted values are the mean values of the third cycle of each concentration. The Tris concentrations of the calibrations were multiplied with the expected desalting level of 98% in order to match the x-axis. (b) Detected apoMb percentage and detected amount of Mb as a function of the feed Tris concentration during the experiments. The values are mean values of the third cycle of the µSMB process for each concentration. (c) Detected apoMb percentage and Tris signal level for three consecutive cycles (4–6) of µSMB run 2 with a constant Tris concentration of 10 mM in the feed stream. The apoMb level was calculated for each switching interval, as indicated by the dotted vertical linesCC BY · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Show all 1 references

Research works printed but not linked: 40

  1. Luis SV, García-Verdugo E. Chemical reactions and processes under flow conditions. Royal Society of Chemistry; 2010.
  2. Gerstweiler L, Bi J, Middelberg APJ. Continuous downstream bioprocessing for intensified manufacture of biopharmaceuticals and antibodies. Chem Eng Sci. 2020. https://doi.org/10.1016/j.ces.2020.116272. · DOI
  3. Rathore AS, Thakur G, Kateja N. Continuous integrated manufacturing for biopharmaceuticals: a new paradigm or an empty promise? Biotechnol Bioeng. 2023;120:333–51. https://doi.org/10.1002/bit.28235. · DOI · Paper page
  4. Xie W, Pedrielli G (2022) From discovery to production: challenges and novel methodologies for next generation biomanufacturing. In: 2022 Winter Simulation Conference (WSC). pp 238–252. · DOI
  5. Rolinger L, Rüdt M, Hubbuch J. A critical review of recent trends, and a future perspective of optical spectroscopy as PAT in biopharmaceutical downstream processing. Anal Bioanal Chem. 2020;412:2047–64. https://doi.org/10.1007/s00216-020-02407-z. · DOI · Paper page
  6. Wasalathanthri DP, Shah R, Ding J, Leone A, Li ZJ. Process analytics 4.0: a paradigm shift in rapid analytics for biologics development. Biotechnol Prog. 2021;37:e3177. https://doi.org/10.1002/btpr.3177. · DOI · Paper page
  7. Lyubarskaya Y, Kobayashi K, Swann P. Application of mass spectrometry to facilitate advanced process controls of biopharmaceutical manufacture. Pharm Bioprocess. 2015;3:313–21. https://doi.org/10.4155/pbp.15.10. · DOI
  8. São Pedro MN, Klijn ME, Eppink MH, Ottens M. Process analytical technique (PAT) miniaturization for monoclonal antibody aggregate detection in continuous downstream processing. J Chem Technol Biotechnol. 2022;97:2347–64. https://doi.org/10.1002/jctb.6920. · DOI
  9. Lingg N, Zhang P, Song Z, Bardor M. The sweet tooth of biopharmaceuticals: importance of recombinant protein glycosylation analysis. Biotechnol J. 2012;7:1462–72. https://doi.org/10.1002/biot.201200078. · DOI
  10. Dotz V, Haselberg R, Shubhakar A, Kozak RP, Falck D, Rombouts Y, Reusch D, Somsen GW, Fernandes DL, Wuhrer M. Mass spectrometry for glycosylation analysis of biopharmaceuticals. TrAC Trends Anal Chem. 2015;73:1–9. https://doi.org/10.1016/j.trac.2015.04.024. · DOI
  11. Füssl F, Cook K, Scheffler K, Farrell A, Mittermayr S, Bones J. Charge variant analysis of monoclonal antibodies using direct coupled pH gradient cation exchange chromatography to high-resolution native mass spectrometry. Anal Chem. 2018;90:4669–76. https://doi.org/10.1021/acs.analchem.7b05241. · DOI
  12. Kükrer B, Filipe V, van Duijn E, Kasper PT, Vreeken RJ, Heck AJR, Jiskoot W. Mass spectrometric analysis of intact human monoclonal antibody aggregates fractionated by size-exclusion chromatography. Pharm Res. 2010;27:2197–204. https://doi.org/10.1007/s11095-010-0224-5. · DOI
  13. Vallejo DD, Jeon CK, Parson KF, Herderschee HR, Eschweiler JD, Filoti DI, Ruotolo BT. Ion mobility–mass spectrometry reveals the structures and stabilities of biotherapeutic antibody aggregates. Anal Chem. 2022;94:6745–53. https://doi.org/10.1021/acs.analchem.2c00160. · DOI
  14. Disela R, Bussy OL, Geldhof G, Pabst M, Ottens M (2023) Characterisation of the E. coli HMS174 and BLR host cell proteome to guide purification process development. Biotechnol J n/a:2300068. https://doi.org/10.1002/biot.202300068. · DOI
  15. Kufer R, Haindl M, Wegele H, Wohlrab S. Evaluation of peptide fractionation and native digestion as two novel sample preparation workflows to improve HCP characterization by LC–MS/MS. Anal Chem. 2019;91:9716–23. https://doi.org/10.1021/acs.analchem.9b01259. · DOI
  16. Yang F, Walker DE, Schoenfelder J, Carver J, Zhang A, Li D, Harris R, Stults JT, Yu XC, Michels DA. A 2D LC-MS/MS strategy for reliable detection of 10-ppm level residual host cell proteins in therapeutic antibodies. Anal Chem. 2018;90:13365–72. https://doi.org/10.1021/acs.analchem.8b03044. · DOI
  17. Wang L, Zeng S, Chen T, Qu H. Direct analysis in real time mass spectrometry, a process analytical technology tool for real-time process monitoring in botanical drug manufacturing. J Pharm Biomed Anal. 2014;91:202–9. https://doi.org/10.1016/j.jpba.2013.12.034. · DOI
  18. Yan B, Chen T, Xu Z, Qu H. Rapid process development of chromatographic process using direct analysis in real time mass spectrometry as a process analytical technology tool. J Pharm Biomed Anal. 2014;94:106–10. https://doi.org/10.1016/j.jpba.2014.01.033. · DOI
  19. Venter A, Nefliu M, Graham Cooks R. Ambient desorption ionization mass spectrometry. TrAC Trends Anal Chem. 2008;27:284–90. https://doi.org/10.1016/j.trac.2008.01.010. · DOI
  20. Gross JH. Direct analysis in real time—a critical review on DART-MS. Anal Bioanal Chem. 2014;406:63–80. https://doi.org/10.1007/s00216-013-7316-0. · DOI
  21. Shieh I-F, Lee C-Y, Shiea J. Eliminating the interferences from TRIS buffer and SDS in protein analysis by fused-droplet electrospray ionization mass spectrometry. J Proteome Res. 2005;4:606–12. https://doi.org/10.1021/pr049765m. · DOI
  22. Chien C-T, Li F-A, Huang J-L, Her G-R. CE-MS of antihistamines using nonvolatile phosphate buffer. Electrophoresis. 2007;28:1454–60. https://doi.org/10.1002/elps.200600542. · DOI
  23. Diehm J, Ballweg T, Franzreb M. Development of a 3D printed micro simulated moving bed chromatography system. J Chromatogr A. 2023;1695:463928. https://doi.org/10.1016/j.chroma.2023.463928. · DOI
  24. Juza M, Mazzotti M, Morbidelli M. Simulated moving-bed chromatography and its application to chirotechnology. Trends Biotechnol. 2000;18:108–18. https://doi.org/10.1016/S0167-7799(99)01419-5. · DOI
  25. Seidel-Morgenstern A, Keßler LC, Kaspereit M. New developments in simulated moving bed chromatography. Chem Eng Technol. 2008;31:826–37. https://doi.org/10.1002/ceat.200800081. · DOI
  26. Mazzotti M, Storti G, Morbidelli M. Optimal operation of simulated moving bed units for nonlinear chromatographic separations. J Chromatogr A. 1997;769:3–24. https://doi.org/10.1016/S0021-9673(97)00048-4. · DOI
  27. Brenner-Weiss G, Kirschhöfer F, Kühl B, Nusser M, Obst U. Analysis of non-covalent protein complexes by capillary electrophoresis–time-of-flight mass spectrometry. J Chromatogr A. 2003;1009:147–53. https://doi.org/10.1016/S0021-9673(03)00553-3. · DOI
  28. Ventouri IK, Malheiro DBA, Voeten RLC, Kok S, Honing M, Somsen GW, Haselberg R. Probing protein denaturation during size-exclusion chromatography using native mass spectrometry. Anal Chem. 2020;92:4292–300. https://doi.org/10.1021/acs.analchem.9b04961. · DOI
  29. Diehm J, Hackert V, Franzreb M. Configurable 3D printed microfluidic multiport valves with axial compression. Micromachines. 2021;12:1247. https://doi.org/10.3390/mi12101247. · DOI
  30. Leweke S, von Lieres E. Chromatography Analysis and Design Toolkit (CADET). Comput Chem Eng. 2018;113:274–94. https://doi.org/10.1016/j.compchemeng.2018.02.025. · DOI
  31. He Q-L, Leweke S, von Lieres E. Efficient numerical simulation of simulated moving bed chromatography with a single-column solver. Comput Chem Eng. 2018;111:183–98. https://doi.org/10.1016/j.compchemeng.2017.12.022. · DOI
  32. Chung SF, Wen CY. Longitudinal dispersion of liquid flowing through fixed and fluidized beds. AIChE J. 1968;14:857–66. https://doi.org/10.1002/aic.690140608. · DOI
  33. Wilson EJ, Geankoplis CJ. Liquid mass transfer at very low Reynolds numbers in packed beds. Ind Eng Chem Fundam. 1966;5:9–14. https://doi.org/10.1021/i160017a002. · DOI
  34. Polson A. The some aspects of diffusion in solution and a definition of a colloidal particle. J Phys Colloid Chem. 1950;54:649–52. https://doi.org/10.1021/j150479a007. · DOI
  35. Schmidt-Traub H, Schulte M, Seidel-Morgenstern A. Preparative chromatography. 2nd ed. John Wiley & Sons, Ltd; 2012. https://doi.org/10.1002/9783527649280. · DOI
  36. Katta V, Chait BT. Observation of the heme-globin complex in native myoglobin by electrospray-ionization mass spectrometry. J Am Chem Soc. 1991;113:8534–5. https://doi.org/10.1021/ja00022a058. · DOI
  37. Zaia J, Annan RS, Biemann K. The correct molecular weight of myoglobin, a common calibrant for mass spectrometry. Rapid Commun Mass Spectrom. 1992;6:32–6. https://doi.org/10.1002/rcm.1290060108. · DOI
  38. Sugiyama K, Highet RJ, Woods A, Cotter RJ, Osawa Y. Hydrogen peroxide-mediated alteration of the heme prosthetic group of metmyoglobin to an iron chlorin product: evidence for a novel oxidative pathway. Proc Natl Acad Sci U S A. 1997;94:796–801. https://doi.org/10.1073/pnas.94.3.796. · DOI
  39. Gomes PS, Zabkova M, Zabka M, Minceva M, Rodrigues AE. Separation of chiral mixtures in real SMB units: the FlexSMB-LSRE®. AIChE J. 2010;56:125–42. https://doi.org/10.1002/aic.11962. · DOI
  40. Beccaria M, Cabooter D. Current developments in LC-MS for pharmaceutical analysis. Analyst. 2020;145:1129–57. https://doi.org/10.1039/C9AN02145K. · DOI

Cited by

Sorted by citations

See all 2

Metadata

Authors
Diehm, J., Witting, L., Kirschhöfer, F., Brenner-Weiß, G., & Franzreb, M.
ORCID
Venue
Analytical and Bioanalytical Chemistry
Publisher
Springer Science and Business Media LLC
Published
2023-11-10
Volume
416
Issue
2
Pages
373-386
ISSN
  • 1618-2642
  • 1618-2650
DOI
10.1007/s00216-023-05023-9
PMC
PMC10761468
Cited by
2
References
1
Language
en
Licence
Funders
  • Stiftung Begabtenförderung Cusanuswerk
  • Karlsruher Institut für Technologie (KIT)

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

How to cite

Cite CoChat for the checks on this page, and the paper for its own findings.

This page
CoChat. (2026). Citation check: Micro simulated moving bed chromatography-mass spectrometry as a continuous on-line process analytical tool. https://app.cochat.ai/papers/10.1007/s00216-023-05023-9
The paper
Diehm, J., Witting, L., Kirschhöfer, F., Brenner-Weiß, G., & Franzreb, M. (2024). Micro simulated moving bed chromatography-mass spectrometry as a continuous on-line process analytical tool. Analytical and Bioanalytical Chemistry. https://doi.org/10.1007/s00216-023-05023-9