CoChat
Sign Up
Cited by 6

From cell clones to recombinant protein product heterogeneity in Chinese hamster ovary cell systems

Wang, Chong, Guo, Xiao, Wang, Wen, Li, Jia-Xin, Wang, Tian-Yun·2025·International Journal of Molecular Sciences
DOI 10.3390/ijms26031324PMC PMC11818180
Open this paper in CoChat — free account

Abstract

Chinese hamster ovary (CHO) cells are commonly used to produce recombinant therapeutic proteins (RTPs). The yield of RTPs in CHO cells has been greatly improved through cell editing and optimization of culture media, cell culture processes, and expression vectors. However, the heterogeneity of cell clones and product aggregation considerably affect the yield and quality of RTPs. Recently, novel technologies such as semi-targeted and site-specific transgene integration, endoplasmic reticulum-residents, and cell culture process optimization have been used to address these issues. In this review, novel developments in the field of CHO cell expression system heterogeneity are summarized. Moreover, the advantages and limitations of the new strategies are discussed, and important methods for the control of RTP quality are outlined.

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 · 10.3390/ijms26031324

Cites

2 of 2 references verified0 citation errors · 0 not checkable

Statistics

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?

  • Recombinant proteins, including growth factors, may exhibit batch-to-batch variability arising from differences in expression systems, host cell biology, post-translational modifications or manufacturing conditions.

    “Recombinant proteins, including growth factors, may also exhibit batch-to-batch variability arising from differences in expression systems, host cell biology, post-translational modifications or manufacturing conditions (O'Flaherty et al., 2020; Wang et al., 2025).”

    From Challenges and solutions in transitioning to animal-free standards: a comprehensive analysis of components in human cell-based developmental neurotoxicity assays (2026) · cites this paper as Wang et al., 2025

    Section 2.3 reports that instability in CHO cell lines can lead to variation in recombinant-protein yield and quality, while Sections 2.4 and 3.2 describe effects of culture conditions and post-translational modifications. The paper focuses on CHO systems; the passages found do not establish batch-to-batch variability specifically, address differences between expression systems, or identify growth factors as an example. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

Self-check

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

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

Cites

Flagged first, then by citations

Research works printed but not linked: 117

  1. [1]Walsh G. Walsh E. Biopharmaceutical benchmarks 2022 Nat. Biotechnol. 2022 40 1722 1760 10.1038/s41587-022-01582-x 36471135 PMC9735008 · DOI
  2. [2]Kim Y.J. Han S.K. Yoon S. Kim C.W. Rich production media as a platform for CHO cell line development AMB Express 2020 10 93 10.1186/s13568-020-01025-3 32415509 PMC7229095 · DOI
  3. [3]Wurm M.J. Wurm F.M. Naming CHO cells for bio-manufacturing: Genome plasticity and variant phenotypes of cell populations in bioreactors question the relevance of old names Biotechnol. J. 2021 16 e2100165 10.1002/biot.202100165 34050613 · DOI
  4. [4]Kim D. Yoon C. Lee G.M. Small molecule epigenetic modulators for enhancing recombinant antibody production in CHO cell cultures Biotechnol. Bioeng. 2022 119 820 831 10.1002/bit.28013 34961935 · DOI
  5. [5]Hou J.J. Hughes B.S. Smede M. Leung K.M. Levine K. Rigby S. Gray P.P. Munro T.P. High-throughput ClonePix FL analysis of mAb-expressing clones using the UCOE expression system N. Biotechnol. 2014 31 214 220 10.1016/j.nbt.2014.02.002 24518824 · DOI
  6. [6]Zeh N. Schmidt M. Schulz P. Fischer S. The new frontier in CHO cell line development: From random to targeted transgene integration technologies Biotechnol. Adv. 2024 75 108402 10.1016/j.biotechadv.2024.108402 38950872 · DOI
  7. [7]Wang Q. Yin B. Chung C.Y. Betenbaugh M.J. Glycoengineering of CHO Cells to Improve Product Quality Methods Mol. Biol. 2017 1603 25 44 10.1007/978-1-4939-6972-2_2 28493121 · DOI
  8. [8]Yang Y. Li Z. Li Q. Ma K. Lin Y. Feng H. Wang T. Increase recombinant antibody yields through optimizing vector design and production process in CHO cells Appl. Microbiol. Biotechnol. 2022 106 4963 4975 10.1007/s00253-022-12051-5 35788878 · DOI
  9. [9]Pilbrough W. Munro T.P. Gray P. Intraclonal protein expression heterogeneity in recombinant CHO cells PLoS ONE 2009 4 e8432 10.1371/journal.pone.0008432 20037651 PMC2793030 · DOI
  10. [10]Wang Q. Aliyu L. Chung C.Y. Rosenberg J.N. Yu G. Betenbaugh M.J. Application of the CRISPR/Cas9 Gene Editing Method for Modulating Antibody Fucosylation in CHO Cells Methods Mol. Biol. 2024 2810 249 271 10.1007/978-1-0716-3878-1_16 38926284 · DOI
  11. [11]Li H. Chen K. Wang Z. Li D. Lin J. Yu C. Yu F. Wang X. Huang L. Jiang C. Genetic analysis of the clonal stability of Chinese hamster ovary cells for recombinant protein production Mol. Biosyst. 2016 12 102 109 10.1039/C5MB00627A 26563441 · DOI
  12. [12]Tihanyi B. Nyitray L. Recent advances in CHO cell line development for recombinant protein production Drug Discov. Today Technol. 2020 38 25 34 10.1016/j.ddtec.2021.02.003 34895638 · DOI
  13. [13]Huhn S.C. Chang M. Jiang B. Tang X. Betenbaugh M. Du Z. Genomic features of recombinant CHO clones arising from transposon-based and randomized integration J. Biotechnol. 2023 373 73 81 10.1016/j.jbiotec.2023.05.009 37271453 · DOI
  14. [14]Lee J.S. Kallehauge T.B. Pedersen L.E. Kildegaard H.F. Site-specific integration in CHO cells mediated by CRISPR/Cas9 and homology-directed DNA repair pathway Sci. Rep. 2015 5 8572 10.1038/srep08572 25712033 PMC4339809 · DOI
  15. [15]Zhang S. Hughes J.D. Murgolo N. Levitan D. Chen J. Liu Z. Shi S. Mutation Detection in an Antibody-Producing Chinese Hamster Ovary Cell Line by Targeted RNA Sequencing Biomed. Res. Int. 2016 2016 8356435 10.1155/2016/8356435 27088091 PMC4818804 · DOI
  16. [16]Barzadd M.M. Lundqvist M. Harris C. Malm M. Volk A.L. Thalén N. Chotteau V. Grassi L. Smith A. Abadi M.L. Autophagy and intracellular product degradation genes identified by systems biology analysis reduce aggregation of bispecific antibody in CHO cells N. Biotechnol. 2022 68 68 76 10.1016/j.nbt.2022.01.010 35123066 · DOI
  17. [17]Srirangan K. Loignon M. Durocher Y. The use of site-specific recombination and cassette exchange technologies for monoclonal antibody production in Chinese Hamster ovary cells: Retrospective analysis and future directions Crit. Rev. Biotechnol. 2020 40 833 851 10.1080/07388551.2020.1768043 32456474 · DOI
  18. [18]Cabaniols J.P. Ouvry C. Lamamy V. Fery I. Craplet M.L. Moulharat N. Guenin S.P. Bedut S. Nosjean O. Ferry G. Meganuclease-driven targeted integration in CHO-K1 cells for the fast generation of HTS-compatible cell-based assays J. Biomol. Screen. 2010 15 956 967 10.1177/1087057110375115 20625180 · DOI · Paper page
  19. [19]Chi X. Zheng Q. Jiang R. Chen-Tsai R.Y. Kong L.J. A system for site-specific integration of transgenes in mammalian cells PLoS ONE 2019 14 e0219842 10.1371/journal.pone.0219842 31344144 PMC6657834 · DOI
  20. [20]Glinšek K. Bozovičar K. Bratkovič T. CRISPR Technologies in Chinese Hamster Ovary Cell Line Engineering Int. J. Mol. Sci. 2023 24 8144 10.3390/ijms24098144 37175850 PMC10179654 · DOI
  21. [21]Bandyopadhyay A.A. O’Brien S.A. Zhao L. Fu H.Y. Vishwanathan N. Hu W.S. Recurring genomic structural variation leads to clonal instability and loss of productivity Biotechnol. Bioeng. 2019 116 41 53 10.1002/bit.26823 30144379 PMC7058117 · DOI
  22. [22]Lee J.S. Kildegaard H.F. Lewis N.E. Lee G.M. Mitigating Clonal Variation in Recombinant Mammalian Cell Lines Trends Biotechnol. 2019 37 931 942 10.1016/j.tibtech.2019.02.007 30898338 · DOI
  23. [23]Sorourian S. Behzad Behbahani A. Forouzanfar M. Jafarinia M. Safari F. Time and Cost-Effective Genome Editing Protocol for Simultaneous Caspase 8 Associated Protein 2 Gene Knock in/out in Chinese Hamster Ovary Cells Using CRISPR-Cas9 System Iran. J. Biotechnol. 2024 22 e3714 10.30498/ijb.2024.398567.3714 38827341 PMC11139449 · DOI
  24. [24]Jusiak B. Jagtap K. Gaidukov L. Duportet X. Bandara K. Chu J. Zhang L. Weiss R. Lu T.K. Comparison of Integrases Identifies Bxb1-GA Mutant as the Most Efficient Site-Specific Integrase System in Mammalian Cells ACS Synth. Biol. 2019 8 16 24 10.1021/acssynbio.8b00089 30609349 · DOI
  25. [25]Kawabe Y. Komatsu S. Komatsu S. Murakami M. Ito A. Sakuma T. Nakamura T. Yamamoto T. Kamihira M. Targeted knock-in of an scFv-Fc antibody gene into the hprt locus of Chinese hamster ovary cells using CRISPR/Cas9 and CRIS-PITCh systems J. Biosci. Bioeng. 2018 125 599 605 10.1016/j.jbiosc.2017.12.003 29295784 · DOI
  26. [26]Liu H. Zhang W. Xie L. Hu Y. Identification of a New Integration Site and Study on Site-Specific Integration in CHO-K1 Cells Pharmaceuticals 2023 17 8 10.3390/ph17010008 38275994 PMC10819697 · DOI
  27. [27]Hamaker N.K. Lee K.H. High-efficiency and multilocus targeted integration in CHO cells using CRISPR-mediated donor nicking and DNA repair inhibitors Biotechnol. Bioeng. 2023 120 2419 2440 10.1002/bit.28393 37039773 PMC10524319 · DOI
  28. [28]Zhao M. Wang J. Luo M. Luo H. Zhao M. Han L. Zhang M. Yang H. Xie Y. Jiang H. Rapid development of stable transgene CHO cell lines by CRISPR/Cas9-mediated site-specific integration into C12orf35 Appl. Microbiol. Biotechnol. 2018 102 6105 6117 10.1007/s00253-018-9021-6 29789882 · DOI
  29. [29]Deaven L.L. Petersen D.F. The chromosomes of CHO, an aneuploid Chinese hamster cell line: G-band, C-band, and autoradiographic analyses Chromosoma 1973 41 129 144 10.1007/BF00319690 4120885 · DOI
  30. [30]Wurm F.M. Hacker D. First CHO genome Nat. Biotechnol. 2011 29 718 720 10.1038/nbt.1943 21822249 · DOI
  31. [31]Dahodwala H. Lee K.H. The fickle CHO: A review of the causes, implications, and potential alleviation of the CHO cell line instability problem Curr. Opin. Biotechnol. 2019 60 128 137 10.1016/j.copbio.2019.01.011 30826670 · DOI
  32. [32]Kaas C.S. Kristensen C. Betenbaugh M.J. Andersen M.R. Sequencing the CHO DXB11 genome reveals regional variations in genomic stability and haploidy BMC Genom. 2015 16 160 10.1186/s12864-015-1391-x 25887056 PMC4359788 · DOI
  33. [33]Wong D.C. Wong K.T. Nissom P.M. Heng C.K. Yap M.G. Targeting early apoptotic genes in batch and fed-batch CHO cell cultures Biotechnol. Bioeng. 2006 95 350 361 10.1002/bit.20871 16894638 · DOI
  34. [34]Cost G.J. Freyvert Y. Vafiadis A. Santiago Y. Miller J.C. Rebar E. Collingwood T.N. Snowden A. Gregory P.D. BAK and BAX deletion using zinc-finger nucleases yields apoptosis-resistant CHO cells Biotechnol. Bioeng. 2010 105 330 340 10.1002/bit.22541 19777580 · DOI
  35. [35]Majors B.S. Chiang G.G. Pederson N.E. Betenbaugh M.J. Directed evolution of mammalian anti-apoptosis proteins by somatic hypermutation Protein Eng. Des. Sel. 2012 25 27 38 10.1093/protein/gzr052 22160868 PMC3276308 · DOI
  36. [36]Figueroa B. Jr. Chen S. Oyler G.A. Hardwick J.M. Betenbaugh M.J. Aven and Bcl-xL enhance protection against apoptosis for mammalian cells exposed to various culture conditions Biotechnol. Bioeng. 2004 85 589 600 10.1002/bit.10913 14966800 · DOI
  37. [37]Doolan P. Meleady P. Barron N. Henry M. Gallagher R. Gammell P. Melville M. Sinacore M. McCarthy K. Leonard M. Microarray and proteomics expression profiling identifies several candidates, including the valosin-containing protein (VCP), involved in regulating high cellular growth rate in production CHO cell lines Biotechnol. Bioeng. 2010 106 42 56 10.1002/bit.22670 20091739 · DOI · Paper page
  38. [38]Jaluria P. Betenbaugh M. Konstantopoulos K. Shiloach J. Enhancement of cell proliferation in various mammalian cell lines by gene insertion of a cyclin-dependent kinase homolog BMC Biotechnol. 2007 7 71 10.1186/1472-6750-7-71 17945021 PMC2164945 · DOI · Paper page
  39. [39]Wang T.Y. Guo X. Expression vector cassette engineering for recombinant therapeutic production in mammalian cell systems Appl. Microbiol. Biotechnol. 2020 104 5673 5688 10.1007/s00253-020-10640-w 32372203 · DOI
  40. [40]Ho S.C. Koh E.Y. Soo B.P. Mariati Chao S.H. Yang Y. Evaluating the use of a CpG free promoter for long-term recombinant protein expression stability in Chinese hamster ovary cells BMC Biotechnol. 2016 16 71 10.1186/s12896-016-0300-y 27756290 PMC5070371 · DOI
  41. [41]Mariati Yeo J.H. Koh E.Y. Ho S.C. Yang Y. Insertion of core CpG island element into human CMV promoter for enhancing recombinant protein expression stability in CHO cells Biotechnol. Prog. 2014 30 523 534 10.1002/btpr.1919 24789630 · DOI
  42. [42]Barnes L.M. Bentley C.M. Dickson A.J. Stability of protein production from recombinant mammalian cells Biotechnol. Bioeng. 2003 81 631 639 10.1002/bit.10517 12529877 · DOI
  43. [43]Bailey L.A. Hatton D. Field R. Dickson A.J. Determination of Chinese hamster ovary cell line stability and recombinant antibody expression during long-term culture Biotechnol. Bioeng. 2012 109 2093 2103 10.1002/bit.24485 22896849 · DOI
  44. [44]Veith N. Ziehr H. MacLeod R.A. Reamon-Buettner S.M. Mechanisms underlying epigenetic and transcriptional heterogeneity in Chinese hamster ovary (CHO) cell lines BMC Biotechnol. 2016 16 6 10.1186/s12896-016-0238-0 26800878 PMC4722726 · DOI
  45. [45]Ritter A. Nuciforo S. Schulze A. Oertli M. Rauschert T. Voedisch B. Geisse S. Jostock T. Laux H. Fam60A plays a role for production stabilities of recombinant CHO cell lines Biotechnol. Bioeng. 2017 114 701 704 10.1002/bit.26181 27617904 · DOI
  46. [46]Majumdar S. Desai R. Hans A. Dandekar P. Jain R. From Efficiency to Yield: Exploring Recent Advances in CHO Cell Line Development for Monoclonal Antibodies Mol. Biotechnol. 2024 67 369 392 10.1007/s12033-024-01060-6 38363529 · DOI
  47. [47]Hernández Bort J.A. Hackl M. Höflmayer H. Jadhav V. Harreither E. Kumar N. Ernst W. Grillari J. Borth N. Dynamic mRNA and miRNA profiling of CHO-K1 suspension cell cultures Biotechnol. J. 2012 7 500 515 10.1002/biot.201100143 21751394 · DOI
  48. [48]Le H. Vishwanathan N. Kantardjieff A. Doo I. Srienc M. Zheng X. Somia N. Hu W.S. Dynamic gene expression for metabolic engineering of mammalian cells in culture Metab. Eng. 2013 20 212 220 10.1016/j.ymben.2013.09.004 24055788 · DOI
  49. [49]Dorai H. Corisdeo S. Ellis D. Kinney C. Chomo M. Hawley-Nelson P. Moore G. Betenbaugh M.J. Ganguly S. Early prediction of instability of Chinese hamster ovary cell lines expressing recombinant antibodies and antibody-fusion proteins Biotechnol. Bioeng. 2012 109 1016 1030 10.1002/bit.24367 22068683 · DOI
  50. [50]Bailey-Kellogg C. Gutiérrez A.H. Moise L. Terry F. Martin W.D. De Groot A.S. CHOPPI: A web tool for the analysis of immunogenicity risk from host cell proteins in CHO-based protein production Biotechnol. Bioeng. 2014 111 2170 2182 10.1002/bit.25286 24888712 PMC4282101 · DOI
  51. [51]Park J.H. Jin J.H. Lim M.S. An H.J. Kim J.W. Lee G.M. Proteomic Analysis of Host Cell Protein Dynamics in the Culture Supernatants of Antibody-Producing CHO Cells Sci. Rep. 2017 7 44246 10.1038/srep44246 28281648 PMC5345005 · DOI
  52. [52]Guo X. Wang C. Wang T.Y. Chromatin-modifying elements for recombinant protein production in mammalian cell systems Crit. Rev. Biotechnol. 2020 40 1035 1043 10.1080/07388551.2020.1805401 32777953 · DOI
  53. [53]Harraghy N. Regamey A. Girod P.A. Mermod N. Identification of a potent MAR element from the mouse genome and assessment of its activity in stable and transient transfections J. Biotechnol. 2011 154 11 20 10.1016/j.jbiotec.2011.04.004 21540066 · DOI · Paper page
  54. [54]Kwaks T.H. Otte A.P. Employing epigenetics to augment the expression of therapeutic proteins in mammalian cells Trends Biotechnol. 2006 24 137 142 10.1016/j.tibtech.2006.01.007 16460822 · DOI
  55. [55]Girod P.A. Nguyen D.Q. Calabrese D. Puttini S. Grandjean M. Martinet D. Regamey A. Saugy D. Beckmann J.S. Bucher P. Genome-wide prediction of matrix attachment regions that increase gene expression in mammalian cells Nat. Methods 2007 4 747 753 10.1038/nmeth1076 17676049 · DOI · Paper page
  56. [56]Wang T.Y. Yang R. Qin C. Wang L. Yang X.J. Enhanced expression of transgene in CHO cells using matrix attachment region Cell Biol. Int. 2008 32 1279 1283 10.1016/j.cellbi.2008.07.014 18703149 · DOI
  57. [57]Ley D. Harraghy N. Le Fourn V. Bire S. Girod P.A. Regamey A. Rouleux-Bonnin F. Bigot Y. Mermod N. MAR elements and transposons for improved transgene integration and expression PLoS ONE 2013 8 e62784 10.1371/journal.pone.0062784 23646143 PMC3640020 · DOI
  58. [58]Sharma N. Hollensen A.K. Bak R.O. Staunstrup N.H. Schrøder L.D. Mikkelsen J.G. The impact of cHS4 insulators on DNA transposon vector mobilization and silencing in retinal pigment epithelium cells PLoS ONE 2012 7 e48421 10.1371/journal.pone.0048421 23110238 PMC3482222 · DOI
  59. [59]Wei M. Mi C.L. Jing C.Q. Wang T.Y. Progress of Transposon Vector System for Production of Recombinant Therapeutic Proteins in Mammalian Cells Front. Bioeng. Biotechnol. 2022 10 879222 10.3389/fbioe.2022.879222 35600890 PMC9114503 · DOI
  60. [60]Neville J.J. Orlando J. Mann K. McCloskey B. Antoniou M.N. Ubiquitous Chromatin-opening Elements (UCOEs): Applications in biomanufacturing and gene therapy Biotechnol. Adv. 2017 35 557 564 10.1016/j.biotechadv.2017.05.004 28528197 · DOI
  61. [61]Moritz B. Becker P.B. Göpfert U. CMV promoter mutants with a reduced propensity to productivity loss in CHO cells Sci. Rep. 2015 5 16952 10.1038/srep16952 26581326 PMC4652263 · DOI
  62. [62]Brown A.J. Sweeney B. Mainwaring D.O. James D.C. Synthetic promoters for CHO cell engineering Biotechnol. Bioeng. 2014 111 1638 1647 10.1002/bit.25227 24615264 · DOI
  63. [63]Liu H.N. Wang X.Y. Zou Y. Wu W.B. Lin Y. Ji B.Y. Wang T.Y. Synthetic enhancers including TFREs improve transgene expression in CHO cells Heliyon 2024 10 e26901 10.1016/j.heliyon.2024.e26901 38468921 PMC10926067 · DOI
  64. [64]Vázquez-Rey M. Lang D.A. Aggregates in monoclonal antibody manufacturing processes Biotechnol. Bioeng. 2011 108 1494 1508 10.1002/bit.23155 21480193 · DOI
  65. [65]Xu T. Zhang J. Wang T. Wang X. Recombinant antibodies aggregation and overcoming strategies in CHO cells Appl. Microbiol. Biotechnol. 2022 106 3913 3922 10.1007/s00253-022-11977-0 35608667 · DOI
  66. [67]Zhou Y. Raju R. Alves C. Gilbert A. Debottlenecking protein secretion and reducing protein aggregation in the cellular host Curr. Opin. Biotechnol. 2018 53 151 157 10.1016/j.copbio.2018.01.007 29414073 · DOI
  67. [68]Andrade C. Arnold L. Motabar D. Aspelund M. Tang A. Hunter A. Chung W.K. An Integrated Approach to Aggregate Control for Therapeutic Bispecific Antibodies Using an Improved Three Column Mab Platform-Like Purification Process Biotechnol. Prog. 2019 35 e2720 10.1002/btpr.2720 30298991 PMC6667909 · DOI
  68. [69]Ratanji K.D. Derrick J.P. Dearman R.J. Kimber I. Immunogenicity of therapeutic proteins: Influence of aggregation J. Immunotoxicol. 2014 11 99 109 10.3109/1547691X.2013.821564 23919460 PMC4002659 · DOI
  69. [70]Rosenberg A.S. Effects of protein aggregates: An immunologic perspective AAPS J. 2006 8 59 10.1208/aapsj080359 17025268 PMC2761057 · DOI · Paper page
  70. [71]Owji H. Nezafat N. Negahdaripour M. Hajiebrahimi A. Ghasemi Y. A comprehensive review of signal peptides: Structure, roles, and applications Eur. J. Cell Biol. 2018 97 422 441 10.1016/j.ejcb.2018.06.003 29958716 · DOI · Paper page
  71. [72]Tripathi N.K. Shrivastava A. Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development Front. Bioeng. Biotechnol. 2019 7 420 10.3389/fbioe.2019.00420 31921823 PMC6932962 · DOI
  72. [73]Mead E.J. Chiverton L.M. Smales C.M. von der Haar T. Identification of the limitations on recombinant gene expression in CHO cell lines with varying luciferase production rates Biotechnol. Bioeng. 2009 102 1593 1602 10.1002/bit.22201 19090535 · DOI
  73. [74]Torres M. Hussain H. Dickson A.J. The secretory pathway—The key for unlocking the potential of Chinese hamster ovary cell factories for manufacturing therapeutic proteins Crit. Rev. Biotechnol. 2023 43 628 645 10.1080/07388551.2022.2047004 35465810 · DOI
  74. [75]Mathias S. Wippermann A. Raab N. Zeh N. Handrick R. Gorr I. Schulz P. Fischer S. Gamer M. Otte K. Unraveling what makes a monoclonal antibody difficult-to-express: From intracellular accumulation to incomplete folding and degradation via ERAD Biotechnol. Bioeng. 2020 117 5 16 10.1002/bit.27196 31631329 · DOI
  75. [76]Tadauchi T. Lam C. Liu L. Zhou Y. Tang D. Louie S. Snedecor B. Misaghi S. Utilizing a regulated targeted integration cell line development approach to systematically investigate what makes an antibody difficult to express Biotechnol. Prog. 2019 35 e2772 10.1002/btpr.2772 30592567 · DOI
  76. [77]Handlogten M.W. Lee-O’Brien A. Roy G. Levitskaya S.V. Venkat R. Singh S. Ahuja S. Intracellular response to process optimization and impact on productivity and product aggregates for a high-titer CHO cell process Biotechnol. Bioeng. 2018 115 126 138 10.1002/bit.26460 28941283 · DOI
  77. [78]Kirimoto Y. Yamano-Adachi N. Koga Y. Omasa T. Effect of co-overexpression of the cargo receptor ERGIC-53/MCFD2 on antibody production and intracellular IgG secretion in recombinant Chinese hamster ovary cells J. Biosci. Bioeng. 2023 136 400 406 10.1016/j.jbiosc.2022.07.002 35963666 · DOI
  78. [79]Zhang Y. Srivastava V. Zhang B. Mammalian cargo receptors for endoplasmic reticulum-to-Golgi transport: Mechanisms and interactions Biochem. Soc. Trans. 2023 51 971 981 10.1042/BST20220713 37334845 PMC10317151 · DOI
  79. [80]Delic M. Göngrich R. Mattanovich D. Gasser B. Engineering of protein folding and secretion-strategies to overcome bottlenecks for efficient production of recombinant proteins Antioxid. Redox Signal. 2014 21 414 437 10.1089/ars.2014.5844 24483278 · DOI
  80. [81]Yamagishi N. Goto K. Nakagawa S. Saito Y. Hatayama T. Hsp105 reduces the protein aggregation and cytotoxicity by expanded-polyglutamine proteins through the induction of Hsp70 Exp. Cell Res. 2010 316 2424 2433 10.1016/j.yexcr.2010.06.003 20542028 · DOI
  81. [82]Hetz C. Zhang K. Kaufman R.J. Mechanisms, regulation and functions of the unfolded protein response Nat. Rev. Mol. Cell Biol. 2020 21 421 438 10.1038/s41580-020-0250-z 32457508 PMC8867924 · DOI · Paper page
  82. [83]Tutar L. Tutar Y. Heat shock proteins; an overview Curr. Pharm. Biotechnol. 2010 11 216 222 10.2174/138920110790909632 20170474 · DOI
  83. [84]Gething M.J. Sambrook J. Protein folding in the cell Nature 1992 355 33 45 10.1038/355033a0 1731198 · DOI
  84. [85]Shamsi T.N. Athar T. Parveen R. Fatima S. A review on protein misfolding, aggregation and strategies to prevent related ailments Int. J. Biol. Macromol. 2017 105 993 1000 10.1016/j.ijbiomac.2017.07.116 28743576 · DOI
  85. [86]Wang W. Roberts C.J. Protein aggregation—Mechanisms, detection, and control Int. J. Pharm. 2018 550 251 268 10.1016/j.ijpharm.2018.08.043 30145245 · DOI
  86. [88]Li W. Prabakaran P. Chen W. Zhu Z. Feng Y. Dimitrov D.S. Antibody Aggregation: Insights from Sequence and Structure Antibodies 2016 5 19 10.3390/antib5030019 31558000 PMC6698864 · DOI
  87. [89]Buchanan A. Clementel V. Woods R. Harn N. Bowen M.A. Mo W. Popovic B. Bishop S.M. Dall’Acqua W. Minter R. Engineering a therapeutic IgG molecule to address cysteinylation, aggregation and enhance thermal stability and expression MAbs 2013 5 255 262 10.4161/mabs.23392 23412563 PMC3893235 · DOI
  88. [90]Bönisch M. Sellmann C. Maresch D. Halbig C. Becker S. Toleikis L. Hock B. Rüker F. Novel CH1:CL interfaces that enhance correct light chain pairing in heterodimeric bispecific antibodies Protein Eng. Des. Sel. 2017 30 685 696 10.1093/protein/gzx044 28981885 PMC5914326 · DOI
  89. [91]Schanzer J.M. Wartha K. Croasdale R. Moser S. Künkele K.P. Ries C. Scheuer W. Duerr H. Pompiati S. Pollman J. A novel glycoengineered bispecific antibody format for targeted inhibition of epidermal growth factor receptor (EGFR) and insulin-like growth factor receptor type I (IGF-1R) demonstrating unique molecular properties J. Biol. Chem. 2014 289 18693 18706 10.1074/jbc.M113.528109 24841203 PMC4081915 · DOI
  90. [92]Bayat H. Hossienzadeh S. Pourmaleki E. Ahani R. Rahimpour A. Evaluation of different vector design strategies for the expression of recombinant monoclonal antibody in CHO cells Prep. Biochem. Biotechnol. 2018 48 160 164 10.1080/10826068.2017.1421966 29313429 · DOI
  91. [93]Eisenhut P. Mebrahtu A. Moradi Barzadd M. Thalén N. Klanert G. Weinguny M. Sandegren A. Su C. Hatton D. Borth N. Systematic use of synthetic 5′-UTR RNA structures to tune protein translation improves yield and quality of complex proteins in mammalian cell factories Nucleic Acids Res. 2020 48 e119 10.1093/nar/gkaa847 33051690 PMC7672427 · DOI · Paper page
  92. [94]Ho S.C. Wang T. Song Z. Yang Y. IgG Aggregation Mechanism for CHO Cell Lines Expressing Excess Heavy Chains Mol. Biotechnol. 2015 57 625 634 10.1007/s12033-015-9852-7 25744439 · DOI
  93. [95]Kober L. Zehe C. Bode J. Optimized signal peptides for the development of high expressing CHO cell lines Biotechnol. Bioeng. 2013 110 1164 1173 10.1002/bit.24776 23124363 · DOI
  94. [96]Le Fourn V. Girod P.A. Buceta M. Regamey A. Mermod N. CHO cell engineering to prevent polypeptide aggregation and improve therapeutic protein secretion Metab. Eng. 2014 21 91 102 10.1016/j.ymben.2012.12.003 23380542 · DOI · Paper page
  95. [97]Galluzzi L. Pietrocola F. Levine B. Kroemer G. Metabolic control of autophagy Cell 2014 159 1263 1276 10.1016/j.cell.2014.11.006 25480292 PMC4500936 · DOI · Paper page
  96. [98]Kaur J. Debnath J. Autophagy at the crossroads of catabolism and anabolism Nat. Rev. Mol. Cell Biol. 2015 16 461 472 10.1038/nrm4024 26177004 · DOI
  97. [99]Boada-Romero E. Letek M. Fleischer A. Pallauf K. Ramón-Barros C. Pimentel-Muiños F.X. TMEM59 defines a novel ATG16L1-binding motif that promotes local activation of LC3 EMBO J. 2013 32 566 582 10.1038/emboj.2013.8 23376921 PMC3579146 · DOI
  98. [100]Wang W. Protein aggregation and its inhibition in biopharmaceutics Int. J. Pharm. 2005 289 1 30 10.1016/j.ijpharm.2004.11.014 15652195 · DOI
  99. [101]Purdie J.L. Kowle R.L. Langland A.L. Patel C.N. Ouyang A. Olson D.J. Cell culture media impact on drug product solution stability Biotechnol. Prog. 2016 32 998 1008 10.1002/btpr.2289 27111574 · DOI
  100. [102]Li Z.M. Fan Z.L. Wang X.Y. Wang T.Y. Factors Affecting the Expression of Recombinant Protein and Improvement Strategies in Chinese Hamster Ovary Cells Front. Bioeng. Biotechnol. 2022 10 880155 10.3389/fbioe.2022.880155 35860329 PMC9289362 · DOI
  101. [103]Zhang W. Liu X. Tang H. Zhang X. Zhou Y. Fan L. Wang H. Tan W.S. Zhao L. Investigation into the impact of tyrosine on the product formation and quality attributes of mAbs in rCHO cell cultures Appl. Microbiol. Biotechnol. 2020 104 6953 6966 10.1007/s00253-020-10744-3 32577803 · DOI
  102. [104]Hwang S.J. Jeon C.J. Cho S.M. Lee G.M. Yoon S.K. Effect of chemical chaperone addition on production and aggregation of recombinant flag-tagged COMP-angiopoietin 1 in Chinese hamster ovary cells Biotechnol. Prog. 2011 27 587 591 10.1002/btpr.579 21425253 · DOI
  103. [105]Onitsuka M. Tatsuzawa M. Asano R. Kumagai I. Shirai A. Maseda H. Omasa T. Trehalose suppresses antibody aggregation during the culture of Chinese hamster ovary cells J. Biosci. Bioeng. 2014 117 632 638 10.1016/j.jbiosc.2013.10.022 24315529 · DOI
  104. [106]Chollet M.E. Skarpen E. Iversen N. Sandset P.M. Skretting G. The chemical chaperone sodium 4-phenylbutyrate improves the secretion of the protein CA267T mutant in CHO-K1 cells trough the GRASP55 pathway Cell Biosci. 2015 5 57 10.1186/s13578-015-0048-4 26457178 PMC4599753 · DOI
  105. [107]Kuwae S. Miyakawa I. Doi T. Development of a chemically defined platform fed-batch culture media for monoclonal antibody-producing CHO cell lines with optimized choline content Cytotechnology 2018 70 939 948 10.1007/s10616-017-0185-1 29322349 PMC6021290 · DOI
  106. [108]Rodriguez J. Spearman M. Tharmalingam T. Sunley K. Lodewyks C. Huzel N. Butler M. High productivity of human recombinant beta-interferon from a low-temperature perfusion culture J. Biotechnol. 2010 150 509 518 10.1016/j.jbiotec.2010.09.959 20933553 · DOI
  107. [109]Gomez N. Wieczorek A. Lu F. Bruno R. Diaz L. Agrawal N.J. Daris K. Culture temperature modulates half antibody and aggregate formation in a Chinese hamster ovary cell line expressing a bispecific antibody Biotechnol. Bioeng. 2018 115 2930 2940 10.1002/bit.26803 30036451 · DOI
  108. [110]Hwang S.J. Yoon S.K. Koh G.Y. Lee G.M. Effects of culture temperature and pH on flag-tagged COMP angiopoietin-1 (FCA1) production from recombinant CHO cells: FCA1 aggregation Appl. Microbiol. Biotechnol. 2011 91 305 315 10.1007/s00253-011-3266-7 21509567 · DOI
  109. [111]Ju H.K. Hwang S.J. Jeon C.J. Lee G.M. Yoon S.K. Use of NaCl prevents aggregation of recombinant COMP-angiopoietin-1 in Chinese hamster ovary cells J. Biotechnol. 2009 143 145 150 10.1016/j.jbiotec.2009.06.017 19559063 · DOI
  110. [112]Han Y.K. Koo T.Y. Lee G.M. Enhanced interferon-beta production by CHO cells through elevated osmolality and reduced culture temperature Biotechnol. Prog. 2009 25 1440 1447 10.1002/btpr.234 19572287 · DOI
  111. [113]Bickel F. Herold E.M. Signes A. Romeijn S. Jiskoot W. Kiefer H. Reversible NaCl-induced aggregation of a monoclonal antibody at low pH: Characterization of aggregates and factors affecting aggregation Eur. J. Pharm. Biopharm. 2016 107 310 320 10.1016/j.ejpb.2016.07.020 27449627 · DOI
  112. [114]Toronjo-Urquiza L. Acosta-Martin A.E. James D.C. Nagy T. Falconer R.J. The use of catechins in Chinese hamster ovary cell media for the improvement of monoclonal antibody yields and a reduction of acidic species Biotechnol. Prog. 2020 36 e2980 10.1002/btpr.2980 32067358 · DOI
  113. [115]Bollati-Fogolín M. Forno G. Nimtz M. Conradt H.S. Etcheverrigaray M. Kratje R. Temperature reduction in cultures of hGM-CSF-expressing CHO cells: Effect on productivity and product quality Biotechnol. Prog. 2005 21 17 21 10.1021/bp049825t 15903236 · DOI
  114. [116]Gomez N. Barkhordarian H. Lull J. Huh J. GhattyVenkataKrishna P. Zhang X. Perfusion CHO cell culture applied to lower aggregation and increase volumetric productivity for a bispecific recombinant protein J. Biotechnol. 2019 304 70 77 10.1016/j.jbiotec.2019.08.001 31381940 · DOI
  115. [117]Sinharoy P. Aziz A.H. Majewska N.I. Ahuja S. Handlogten M.W. Perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells Sci. Rep. 2020 10 16620 10.1038/s41598-020-73573-4 33024175 PMC7538420 · DOI
  116. [118]Rezazade Bazaz M. Ghahramani Seno M.M. Dehghani H. Transposase-CRISPR mediated targeted integration (TransCRISTI) in the human genome Sci. Rep. 2022 12 3390 10.1038/s41598-022-07158-8 35232993 PMC8888626 · DOI
  117. [119]Bire S. Dusserre Y. Bigot Y. Mermod N. PiggyBac transposase and transposon derivatives for gene transfer targeting the ribosomal DNA loci of CHO cells J. Biotechnol. 2021 341 103 112 10.1016/j.jbiotec.2021.09.011 34560160 · DOI

Cited by

Sorted by citations

Authors
Wang, C., Guo, X., Wang, W., Li, J.-X., & Wang, T.-Y.
Affiliations
  • School of Medical Engineering, Xinxiang Medical University, Xinxiang 453003, China
  • School of Life Science and Technology, Xinxiang Medical University, Xinxiang 453003, China
  • International Joint Laboratory of Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang 453003, China
ORCID
Venue
International Journal of Molecular Sciences
Publisher
MDPI AG
Published
2025-02-04
Volume
26
Issue
3
Pages
1324
ISSN
1422-0067
DOI
10.3390/ijms26031324
PMC
PMC11818180
Cited by
6
References
2
Language
en
Licence
creativecommons.org/licenses/by/4.0/ (vor)
Funders
National Natural Science Foundation of China (U23A20270)

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