Protein separation by capillary gel electrophoresis: A review
Open accessAbstract
Capillary gel electrophoresis (CGE) has been used for protein separation for more than two decades. Due to the technology advancement, current CGE methods are becoming more and more robust and reliable for protein analysis, and some of the methods have been routinely used for the analysis of protein-based pharmaceuticals and quality controls. In light of this progress, we survey 147 papers related to CGE separations of proteins and present an overview of this technology. We first introduce briefly the early development of CGE. We then review the methodology, in which we specifically describe the matrices, coatings, and detection strategies used in CGE. CGE using microfabricated channels and incorporation of CGE with two-dimensional protein separations are also discussed in this section. We finally present a few representative applications of CGE for separating proteins in real-world samples.
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Genetics of seed quality in soybean analysed by capillary gel electrophoresis
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Separation of Rubisco in Extracts of Plant Leaves by Capillary Electrophoresis with Sieving Polymers
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Coronas of micro/nano plastics: a key determinant in their risk assessments
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Origin of nanostructural inhomogeneity in polymer-network gels
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Capillary electrophoresis and isoelectric focusing in peptide and protein analysis
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“[121], to which the readers are referred for further insight on these matters.”
BackgroundUV‐Triggered Polymerization, Deposition, and Patterning of Plant Phenolic Compounds
Behboodi‐Sadabad, F., Zhang, H., Trouillet, V., Welle, A., Plumeré, N., & Levkin, P. A. (2017) · Advanced Functional Materials · doi:10.1002/adfm.201700127 · Cited by 158 · see citing works
Thompson, B. L., Ouyang, Y., Duarte, G. R. M., Carrilho, E., Krauss, S. T., & Landers, J. P. (2015) · Nature Protocols · doi:10.1038/nprot.2015.051 · Cited by 149 · see citing works
Recent advances in protein analysis by capillary and microchip electrophoresis
Dawod, M., Arvin, N. E., & Kennedy, R. T. (2017) · The Analyst · doi:10.1039/c7an00198c · Cited by 132 · see citing works
Acoustic Microfluidic Separation Techniques and Bioapplications: A Review
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Microfluidic Isolation and Enrichment of Nanoparticles
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Scattering perspectives on nanostructural inhomogeneity in polymer network gels
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Recent applications of capillary electromigration methods to separation and analysis of proteins
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“aca slightly branched polymers, such as linear polyacrylamide, poly(ethylene oxide), dextran, pullulan and cross-linked polyacrylamide [8].”
BackgroundRecent trends of capillary electrophoresis‐mass spectrometry in proteomics research
Gomes, F. P., & Yates, J. R. (2019) · Mass Spectrometry Reviews · doi:10.1002/mas.21599 · Cited by 112 · see citing works
Recent applications of microchip electrophoresis to biomedical analysis
Nuchtavorn, N., Suntornsuk, W., Lunte, S. M., et al. (2015) · Journal of Pharmaceutical and Biomedical Analysis · doi:10.1016/j.jpba.2015.03.002 · Cited by 109 · see citing works
“proteins, nucleic acids and nanoparticles) [56-59].”
Gahoual, R., Beck, A., Leize‐Wagner, E., & François, Y. (2016) · Journal of Chromatography B · doi:10.1016/j.jchromb.2016.05.028 · Cited by 103 · see citing works
“As the main handicap of CE is known to be the poor UV-detection limit, CE is fully suitable for coupling to different very sensitive detection methods such as laser induced by fluorescence (LIF) [19] and MS [21, 22].”
MethodRecent advances in the analysis of therapeutic proteins by capillary and microchip electrophoresis
Creamer, J. S., Oborny, N. J., & Lunte, S. M. (2014) · Analytical Methods · doi:10.1039/c4ay00447g · Cited by 91 · see citing works
Tetala, K. K. R., & Vijayalakshmi, M. A. (2015) · Analytica Chimica Acta · doi:10.1016/j.aca.2015.11.037 · Cited by 90 · see citing works
Beck, A., D’Atri, V., Ehkirch, A., Fekete, S., Hernandez‐Alba, O., Gahoual, R., Leize‐Wagner, E., François, Y., Guillarme, D., & Cianférani, S. (2019) · Expert Review of Proteomics · doi:10.1080/14789450.2019.1578215 · Cited by 90 · see citing works
“CE-SDS is a method based on the same mechanism as sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE).”
MethodInfluentialMiddle-down approach: a choice to sequence and characterize proteins/proteomes by mass spectrometry
Pandeswari, P. B., & Sabareesh, V. (2019) · RSC Advances · doi:10.1039/c8ra07200k · Cited by 86 · see citing works
Dahmardeh, M., Dastjerdi, H. M., Mazal, H., et al. (2023) · Nature Methods · doi:10.1038/s41592-023-01778-2 · Cited by 86 · see citing works
D’Atri, V., Imiołek, M., Quinn, C. F., Finny, A. S., Lauber, M. A., Fekete, S., & Guillarme, D. (2024) · Journal of Chromatography A · doi:10.1016/j.chroma.2024.464862 · Cited by 77 · see citing works
Lyzenga, W. J., Harrington, M., Bekkaoui, D. R., Wigness, M., Hegedus, D. D., & Rozwadowski, K. L. (2019) · BMC Plant Biology · doi:10.1186/s12870-019-1873-0 · Cited by 74 · see citing works
Peptide-based therapeutics: quality specifications, regulatory considerations, and prospects
Rastogi, S., Shukla, S., Kalaivani, M., & Singh, G. N. (2018) · Drug Discovery Today · doi:10.1016/j.drudis.2018.10.002 · Cited by 72 · see citing works
Palma‐Cando, A., Woitassek, D., Brunklaus, G., & Scherf, U. (2016) · Materials Chemistry Frontiers · doi:10.1039/c6qm00281a · Cited by 69 · see citing works
Invited Article: Terahertz microfluidic chips sensitivity-enhanced with a few arrays of meta-atoms
Serita, K., Matsuda, E., Okada, K., Murakami, H., Kawayama, I., & Tonouchi, M. (2018) · APL Photonics · doi:10.1063/1.5007681 · Cited by 70 · see citing works
Tamizi, E., & Jouyban, A. (2014) · Electrophoresis · doi:10.1002/elps.201400343 · Cited by 68 · see citing works
Antibody validation for Western blot: By the user, for the user
Pillai‐Kastoori, L., Heaton, S., Shiflett, S. D., Roberts, A. C., Solache, A., & Schutz-Geschwender, A. R. (2019) · Journal of Biological Chemistry · doi:10.1074/jbc.ra119.010472 · Cited by 59 · see citing works
“Capillary Western blotting through capillary gel electrophoresis (CGE) replaces the existing gel and membrane system with protein separation, immobilization, and staining in a single column (69).”
MethodDynamics-based assessment of nanoscopic polymer-network mesh structures and their defects
Saalwächter, K., & Seiffert, S. (2018) · Soft Matter · doi:10.1039/c7sm02444d · Cited by 63 · see citing works
Analysis of proteins and peptides by electromigration methods in microchips
Štěpánová, S., & Kašička, V. (2016) · Journal of Separation Science · doi:10.1002/jssc.201600962 · Cited by 62 · see citing works
Antibody validation for Western blot: By the user, for the user
Pillai‐Kastoori, L., Heaton, S., Shiflett, S. D., Roberts, A. C., Solache, A., & Schutz-Geschwender, A. R. (2020) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)49905-4 · Cited by 65 · see citing works
Serita, K., Murakami, H., Kawayama, I., & Tonouchi, M. (2019) · Photonics · doi:10.3390/photonics6010012 · Cited by 61 · see citing works
Current advances and challenges in microfluidic free-flow electrophoresis—A critical review
Novo, P., & Janasek, D. (2017) · Analytica Chimica Acta · doi:10.1016/j.aca.2017.08.017 · Cited by 56 · see citing works
Analysis and applications of nanoparticles in capillary electrophoresis
Ban, E., Yoo, Y. S., & Song, E. J. (2015) · Talanta · doi:10.1016/j.talanta.2015.03.020 · Cited by 54 · see citing works
Analytical aspects of biosimilarity issues of protein drugs
Kálmán-Szekeres, Z., Olajos, M., & Ganzler, K. (2012) · Journal of Pharmaceutical and Biomedical Analysis · doi:10.1016/j.jpba.2012.04.037 · Cited by 53 · see citing works
Ouyang, Y., Wang, S., Li, J., Riehl, P. S., Begley, M. R., & Landers, J. P. (2013) · Lab on a Chip · doi:10.1039/c3lc41275j · Cited by 51 · see citing works
Capillary Gel Electrophoresis of Proteins: Historical overview and recent advances
Hajba, L., Jeong, S., Chung, D. S., et al. (2023) · TrAC Trends in Analytical Chemistry · doi:10.1016/j.trac.2023.117024 · Cited by 51 · see citing works
Polymer sieving matrices in microanalytical electrophoresis
Chung, M., Kim, D., & Herr, A. E. (2014) · The Analyst · doi:10.1039/c4an01179a · Cited by 51 · see citing works
Updated but not outdated “Gliadin”: A plant protein in advanced pharmaceutical nanotechnologies
Mehanna, M. M., & Mneimneh, A. T. (2020) · International Journal of Pharmaceutics · doi:10.1016/j.ijpharm.2020.119672 · Cited by 49 · see citing works
Dadouch, M., Ladner, Y., & Perrin, C. (2021) · Separations · doi:10.3390/separations8010004 · Cited by 50 · see citing works
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Regulatory Guidelines for the Analysis of Therapeutic Peptides and Proteins
Elsayed, Y. Y., Kühl, T., & Imhof, D. (2025) · Journal of Peptide Science · doi:10.1002/psc.70001 · Cited by 47 · see citing works
“…gel electrophoresis (SDS-PAGE) [66], native PAGE [67], isoelectric focusing (IEF) [68], and capillary electrophoresis (CE) [69–71], are commonly used for the quality con-trol of proteins and, due to the sequence length and technical issues, to a much lesser extent of peptides.”
Wang, L., Feng, F., & Ma, Z. (2015) · Scientific Reports · doi:10.1038/srep16855 · Cited by 44 · see citing works
Chowdhury, A. D., Agnihotri, N., Doong, R., et al. (2017) · Analytical Chemistry · doi:10.1021/acs.analchem.7b03095 · Cited by 43 · see citing works
Коноплев, Г., Agafonova, D. S., Bakhchova, L., Mukhin, N. V., Kurachkina, M., Schmidt, M.-P., Верлов, Н. А., Sidorov, A. I., Oseev, A., Stepanova, O. S., et al. (2022) · Biomedicines · doi:10.3390/biomedicines10020207 · Cited by 43 · see citing works
Smith, M. T., Zhang, S., Adams, T., et al. (2017) · Electrophoresis · doi:10.1002/elps.201600519 · Cited by 40 · see citing works
Zhang, C.-X., & Meagher, M. M. (2017) · Analytical Chemistry · doi:10.1021/acs.analchem.6b02933 · Cited by 39 · see citing works
Zhang, Z., Park, J., Barrett, H., Dooley, S., Davies, C., & Verhagen, M. F. J. M. (2020) · Human Gene Therapy · doi:10.1089/hum.2020.233 · Cited by 35 · see citing works
“9 10–12 CE-SDS is now commonly used to determine the protein purity of therapeutic proteins, especially mAbs, due to the simpli-fied sample preparation and direct ultraviolet (UV) detection.”
Emerging Nanoporous Materials for Biomolecule Separation
Song, Y., Bao, H., Shen, X., Li, X., Liang, X., & Wang, S. (2022) · Advanced Functional Materials · doi:10.1002/adfm.202113153 · Cited by 36 · see citing works
Stutz, H. (2022) · Journal of Pharmaceutical and Biomedical Analysis · doi:10.1016/j.jpba.2022.115089 · Cited by 33 · see citing works
“Further details are provided elsewhere [258].”
BackgroundMicrofluidic-based electrically driven particle manipulation techniques for biomedical applications
Wang, J., Cui, X., Wang, W., Wang, J., Zhang, Q. W., Guo, X., Liang, Y., Lin, S., Chu, B., & Cui, D. (2025) · RSC Advances · doi:10.1039/d4ra05571c · Cited by 32 · see citing works
Latosińska, A., Frantzi, M., Vlahou, A., & Mischak, H. (2013) · PROTEOMICS - CLINICAL APPLICATIONS · doi:10.1002/prca.201300038 · Cited by 31 · see citing works
“Several capillary electro-separation modes are available, such as CZE, CIEF [9], capillary gel electrophoresis [10], capillary isotachophoresis [11], micellar electrokinetic capillary chromatography [12], microemulsion electrokinetic chromatography [13], nonaqueous CE [14], and capillary electrochromatography [15].”
MethodCapillary gel electrophoresis for precise protein quantitation
Cianciulli, C., Hahne, T., & Wätzig, H. (2012) · Electrophoresis · doi:10.1002/elps.201200177 · Cited by 31 · see citing works
Esterman, A. L., Katiyar, A., & Krishnamurthy, G. (2016) · Journal of Pharmaceutical and Biomedical Analysis · doi:10.1016/j.jpba.2016.06.006 · Cited by 31 · see citing works
de Oliveira, M. A. L., Porto, B. L. S., Bastos, C. D. A., Sabarense, C. M., Vaz, F. A. S., Neves, L. N. D. O., Duarte, L. M., Campos, N. D. S., Chellini, P. R., da Silva, P. H. F., et al. (2016) · Analytical Methods · doi:10.1039/c5ay02736e · Cited by 30 · see citing works
Sánchez‐Hernández, L., Montealegre, C., Kiessig, S., Moritz, B., & Neusüß, C. (2017) · ELECTROPHORESIS · doi:10.1002/elps.201600464 · Cited by 29 · see citing works
Schlecht, J., Jooß, K., & Neusüß, C. (2018) · Analytical and Bioanalytical Chemistry · doi:10.1007/s00216-018-1157-9 · Cited by 29 · see citing works
Wang, X., An, J., Li, J., et al. (2017) · Microchimica Acta · doi:10.1007/s00604-017-2131-5 · Cited by 28 · see citing works
“Many separation modes have been used in CE, such as capillary zone electrophoresis (CZE) [9, 10], capillary gel electrophoresis (CGE) [11, 12], micellar electrokinetic chromatography (MEKC) [13, 14] and capi l lary electrochromatography (CEC) [15, 16].”
MethodCamperi, J., Cock, B. D., Pichon, V., et al. (2018) · Talanta · doi:10.1016/j.talanta.2018.09.095 · Cited by 28 · see citing works
Recent advances in capillary gel electrophoresis for the analysis of proteins
Bhimwal, R., Rustandi, R. R., Payne, A. F., & Dawod, M. (2022) · Journal of Chromatography A · doi:10.1016/j.chroma.2022.463453 · Cited by 27 · see citing works
van Tricht, E., Geurink, L., Pajic, B., et al. (2015) · Talanta · doi:10.1016/j.talanta.2015.07.047 · Cited by 26 · see citing works
Ecotoxicological significance of bio-corona formation on micro/nanoplastics in aquatic organisms
Camil Rex, M., Debroy, A., Nirmala, M. J., & Mukherjee, A. (2023) · RSC Advances · doi:10.1039/d3ra04054b · Cited by 25 · see citing works
Engineering death resistance in CHO cells for improved perfusion culture
MacDonald, M. A., Nöbel, M., Martínez, V. S., Baker, K. N., Shave, E., Gray, P. P., Mahler, S. M., Munro, T. P., Nielsen, L. K., & Marcellin, E. (2022) · mAbs · doi:10.1080/19420862.2022.2083465 · Cited by 24 · see citing works
“Following the same principles of standard SDS-gel electrophoresis, the capillary setup allows for more reproducible quantification.(45) Regarding perfusion cultures, supernatant and permeate samples on day 17 (end of steady-state) and the end of the run were analyzed.”
MethodMolecularly imprinted polyaniline for detection of horseradish peroxidase
Pidenko, P. S., Pidenko, S. A., Skibina, J. S., et al. (2020) · Analytical and Bioanalytical Chemistry · doi:10.1007/s00216-020-02689-3 · Cited by 24 · see citing works
“It is therefore not surprising that rapid increase in the development of highly efficient methods of protein purification, characterization, quantitation, detection [3, 4], and separation techniques [5, 6] has been seen.”
MethodKubota, K., Kobayashi, N., Yabuta, M., et al. (2017) · Journal of Pharmaceutical and Biomedical Analysis · doi:10.1016/j.jpba.2017.03.027 · Cited by 24 · see citing works
Recent advances in microscale western blotting
Sanders, B. J., Kim, D. C., & Dunn, R. C. (2016) · Analytical Methods · doi:10.1039/c6ay01947a · Cited by 24 · see citing works
High-Performance Polymers for Separation and Purification Processes: An Overview
Khodakarami, M., & Alagha, L. (2017) · Polymer-Plastics Technology and Engineering · doi:10.1080/03602559.2017.1298800 · Cited by 23 · see citing works
Analysis of myofibrillar and sarcoplasmic proteins in pork meat by capillary gel electrophoresis
Grujić, R., Grujić, R., Savanović, D., et al. (2018) · Foods and raw materials · doi:10.21603/2308-4057-2018-2-421-428 · Cited by 23 · see citing works
Duhamel, L., Gu, Y. H., Barnett, G. V., Tao, Y., Voronov, S., Ding, J. H., Mussa, N. A., & Li, Z. J. (2019) · Analytical and Bioanalytical Chemistry · doi:10.1007/s00216-019-01942-8 · Cited by 22 · see citing works
Kim, D. C., & Dunn, R. C. (2015) · Analytical Chemistry · doi:10.1021/acs.analchem.5b04187 · Cited by 21 · see citing works
Nonlinear electrophoretic velocity of a spherical colloidal particle
Cobos, R., & Khair, A. S. (2023) · Journal of Fluid Mechanics · doi:10.1017/jfm.2023.537 · Cited by 20 · see citing works
Protein Analysis-on-Chip Systems in Foodomics
Nazzaro, F., Orlando, P., Fratianni, F., et al. (2012) · Nutrients · doi:10.3390/nu4101475 · Cited by 20 · see citing works
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Evaluation of different analytical methods to assess failure of passive transfer in neonatal calves
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Fabrication of an Open Microfluidic Device for Immunoblotting
Abdel-Sayed, P., Yamauchi, K. A., Gerver, R. E., et al. (2017) · Analytical Chemistry · doi:10.1021/acs.analchem.7b02406 · Cited by 18 · see citing works
Moreno-Gordaliza, E., Stigter, E., Lindenburg, P., & Hankemeier, T. (2016) · Analytica Chimica Acta · doi:10.1016/j.aca.2016.04.005 · Cited by 14 · see citing works
Guzman, N., & Guzman, D. E. (2021) · Biomolecules · doi:10.3390/biom11101443 · Cited by 17 · see citing works
Application of Capillary Electromigration Methods for Physicochemical Measurements
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Covalent Attachment and Detachment by Reactive DESI of Sequence‐Coded Polymer Taggants
Youssef, I., Sergent, I., Konishcheva, E. V., Kebe, S. I., Greff, V., Karamessini, D., Matloubi, M., Ouahabi, A. A., Moesslein, J., Amalian, J., et al. (2022) · Macromolecular Rapid Communications · doi:10.1002/marc.202200412 · Cited by 16 · see citing works
Chevrot, G., Fileti, E. E., & Chaban, V. V. (2016) · Journal of Molecular Modeling · doi:10.1007/s00894-016-3123-9 · Cited by 16 · see citing works
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Napp, A., Houbart, V., Demelenne, A., Merville, M., Crommen, J., Dumoulin, M., Garraux, G., Servais, A., & Fillet, M. (2018) · Electrophoresis · doi:10.1002/elps.201800224 · Cited by 15 · see citing works
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- Authors
- Zhu, Z., Lu, J. J., & Liu, S.
- Affiliations
- University of Oklahoma (US)
- Corresponding author
- Liu, Shaorong
- Venue
- Analytica Chimica Acta
- Publisher
- Elsevier BV
- Published
- 2012-01
- Volume
- 709
- Pages
- 21-31
- ISSN
- 0003-2670 (ISSN-L)
- PMC
- PMC3227876
- OpenAlex
- W2039108689
- Cited by
- 202
- References
- 151
- Citations by year
- 2026: 7 · 2025: 14 · 2024: 9 · 2023: 11 · 2022: 20 · 2021: 9 · 2020: 23 · 2019: 16 · 2018: 14 · 2017: 20 · 2016: 21 · 2015: 16 · 2014: 8 · 2013: 6 · 2012: 8
- FWCI
- 0.57
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Analytica Chimica Acta (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Science Foundation (CHE 1011957)
- National Institutes of Health (RO1 GM078592)
- Oklahoma Center for the Advancement of Science and Technology
- U.S. Department of Energy (SC0006351)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Analytical Chemistry and Chromatography
- Keywords
- capillary gel electrophoresis, protein separation, capillary coating, detection methods, microfabricated channels, quality control
- MeSH terms
- Proteins, Spectrophotometry, Ultraviolet, Immunoblotting, Electrophoresis, Gel, Two-Dimensional, Electrophoresis, Capillary
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