SpringerLink
Log in

Rapid Purification of Immunoglobulin G Using a Protein A-immobilized Monolithic Spin Column with Hydrophilic Polymers

  • Original Paper
  • Published:
Analytical Sciences Aims and scope Submit manuscript

Abstract

A rapid purification method was developed for antibody production in Chinese hamster ovary (CHO) cells using a Protein A-immobilized monolithic silica spin column with hydrophilic polymers. Monolithic silica modified with copolymers of 2-hydroxyethylmethacrylate (HEMA) and glycidyl methacrylate (GMA) showed lower non-specific protein absorption than that modified with a silane reagent. The epoxy group of GMA was converted to an amino group, and Protein A was modified by the coupling reagent. The amount of immobilized Protein A was controlled by changing the ratio of GMA to HEMA and the mesopore size of monolith. A modified monolith disk was fixed to a spin column for rapid antibody purification. The linear curves (for the antibody concentrations over 10 – 300 μg/mL) had a correlation coefficient of >0.999. Our column had various analytical advantages over previously reported columns, including a shorter preparation time (<10 min) and smaller sample volumes for purification with Protein A-immobilized agarose.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. M. Grom, M. Kozorog, S. Caserman, A. Pohar, and B. Likozar, J. Chromatogr. B, 2018, 1083, 44.

    Article  CAS  Google Scholar 

  2. L. M. Barnes, C. M. Bentley, and A. J. Dickson, Cytotechnology, 2000, 32, 109.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. H. Dorai, S. Corisdeo, D. Ellis, C. Kinney, M. Chomo, P. Hawley-Nelson, G. Moore, H. Betenbaugh, and S. Ganguly, Biotechnol. Bioeng., 2012, 109, 1016.

    Article  CAS  PubMed  Google Scholar 

  4. S. Dharshanan, H. Chong, C. S. Hung, Z. Zamrod, and N. Kamal, Electron. J. Biotechnol., 2011, 14, 1.

    Article  Google Scholar 

  5. S. Dharshanan and C. S. Hung, Methods Mol. Biol., 2014, 1131, 105.

    Article  CAS  PubMed  Google Scholar 

  6. B. Zang, J. Ren, L. Xu, and L. Jia, J. Chromatogr. B, 2016, 1008, 132.

    Article  CAS  Google Scholar 

  7. X. Zhang, Y. Duan, and X. Zeng, Process Biochem., 2020, 91, 90.

    Article  CAS  Google Scholar 

  8. X. Zhang, Y. Duan, and X. Zeng, ACS Omega, 2017, 2, 1731.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. A. Denizli, A. Y. Rad, and E. Piskin, J. Chromatogr. B, 1995, 668, 13.

    Article  CAS  Google Scholar 

  10. M. Lorenz, C. Paganini, G. Storti, and M. Morbidelli, Materials, 2019, 12, 1580.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. S. Ohlson and J. Wieslander, J. Chromatogr. A, 1987, 397, 207.

    Article  CAS  Google Scholar 

  12. S. Huang, S. Y. Cheng, S. Y. Zhang, Y. L. Yan, S. L. Cai, X. L. Li, S. R. Zheng, J. Fan, and W. G. Zhang, New J. Chem., 2020, 44, 7884.

    Article  CAS  Google Scholar 

  13. B. N. P. Sah, J. Lueangsakulthai, B. R. Hauser, V. D. Mathieu, B. Scottoline, M. K. Pastey, and D. C. Dallas, Front. Nutr., 2020, 7, 136.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Y. T. Chang, M. C. Chang, Y. J. Tsai, C. Ferng, H. C. Shih, Y. P. Kuo, C. H. Chen, and I. L. Tsai, J. Food Drug Anal., 2019, 27, 475.

    Article  CAS  PubMed  Google Scholar 

  15. E. Lalli, J. S. Silva, C. Boi, and G. C. Sarti, Membranes, 2020, 10, 1.

    Article  CAS  Google Scholar 

  16. U. Černigoj, U. Vidic, B. Nemec, J. Gašperšič, J. Vidič, N. L. Krajnc, A. Štrancar, and A. Podgornik, J. Chromatogr. A, 2016, 1464, 72.

    Article  PubMed  Google Scholar 

  17. M. V. Volokitina, V. A. Korzhikov-Vlakh, T. B. Tennikova, and E. G. Korzhikova-Vlakh, J. Pharm. Biomed. Anal., 2017, 145, 169

    Google Scholar 

  18. M. W. H. Roberts, C. M. Ongkudon, G. M. Forde, and M. K. Danquah, J. Sep. Sci., 2009, 32, 2485.

    Article  CAS  PubMed  Google Scholar 

  19. A. Podgomik, M. Hamachi, Y. Isakari, N. Yoshimoto, and S. Yamamoto, Electrophoresis, 2017, 38, 2892.

    Article  Google Scholar 

  20. H. Minakuchi, K. Nakanishi, N. Soga, N. Ishizuka, and N. Tanaka, J. Chromatogr. A, 1997, 762, 135.

    Article  CAS  PubMed  Google Scholar 

  21. H. Minakuchi, K. Nakanishi, N. Soga, N. Ishizuka, and N. Tanaka, J. Chromatogr. A, 1998, 797, 121.

    Article  CAS  Google Scholar 

  22. A. Namera, A. Nakamoto, M. Nishida, T. Saito, I. Kishiyama, S. Miyazaki, M. Yahata, M. Yashiki, and M. Nagao, J. Chromatogr. A, 2008, 797, 121.

    Google Scholar 

  23. T. Saito, R. Yamamoto, S. Inoue, I. Kishiyama, S. Miyazaki, A. Nakamoto, M. Nishida, A. Namera, and S. Inokuchi, J. Chromatogr. B, 2008, 867, 99.

    Article  CAS  Google Scholar 

  24. P. Kovarik, R. J. Hodgson, T. Covey, M. A. Brook, and J. D. Brenna, Anal. Chem., 2005, 773, 340.

    Google Scholar 

  25. R Mallik, H. Xuan, and D. S. Hage, J. Chromatogr. A, 2007, 1149, 294.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. X. Liu, J. **ng, Y. Guan, G. Shan, and H. Liu, Colloids Surf., A, 2004, 238, 127.

    Article  CAS  Google Scholar 

  27. S. C. Crowley, K. C. Chan, and R. R. Walters, J. Chromatogr. A, 1986, 359, 359.

    Article  CAS  Google Scholar 

  28. T. L. Lasseter, B. H. Clare, N. L. Abbott, and R. J. Hamers, J. Am. Chem. Soc., 2004, 126, 10220.

    Article  CAS  PubMed  Google Scholar 

  29. L. S. Wang, L. C. Wu, S. Y. Lu, L. L. Chang, I. T. Teng, C. M. Yang, and J. A. Ho, ACS Nano, 2010, 8, 4371.

    Article  Google Scholar 

  30. Z. Zhao, A. Malik, and M. L. Lee, Anal. Chem., 1993, 65, 2747.

    Article  CAS  PubMed  Google Scholar 

  31. M. Malmsten and J. M. Van Alstine, J. Colloid Interface Sci., 1996, 177, 502.

    Article  CAS  Google Scholar 

  32. D. Wu, B. Zhao, Z. Dai, J. Qin, and B. Lin, Lab Chip, 2006, 6, 942.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This research was partially supported by the develo** key technologies for discovering and manufacturing pharmaceuticals used for next-generation treatments and diagnoses both from the Ministry of Economy, Trade and Industry, Japan (METI) and

Author information

Authors and Affiliations

  1. Bio-Process Engineering Laboratory, Graduate School of Yamaguchi University Biomedical Engineering Center (YUBEC), 2-16-1 Tokiwadai, Ube, 755-8611, Japan

    Shigenori Ota, Noriko Yoshimoto & Shuichi Yamamoto

  2. GL Sciences Inc., 237-2 Sayamagahara, Iruma, Saitama, 358-0032, Japan

    Yuko Yui & Tsutomu Sato

Authors
  1. Shigenori Ota
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuko Yui
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tsutomu Sato
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Noriko Yoshimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shuichi Yamamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shigenori Ota.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ota, S., Yui, Y., Sato, T. et al. Rapid Purification of Immunoglobulin G Using a Protein A-immobilized Monolithic Spin Column with Hydrophilic Polymers. ANAL. SCI. 37, 985–990 (2021). https://doi.org/10.2116/analsci.20P378

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2116/analsci.20P378

Keywords

Search

Navigation