Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants

HMM Tariq Hossain; Kwan Yong Choi

doi:doi:10.11648/j.cbe.20180301.11

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Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants

HMM Tariq Hossain, Kwan Yong Choi

Published in Chemical and Biomolecular Engineering (Volume 3, Issue 1)

Received: 31 January 2017 Accepted: 7 December 2017 Published: 2 January 2018

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Abstract

Apart from tradition, the biospecific affinity isolation has become one of the most rapidly growing concerns with cellulose binding domains (CBDs), a high-capacity tag for cost-effective purifications of fusion proteins. We report here a new strategy optimized for isolation of two fusion proteins, the FGF-1 (a human functional protein) and the H5N1 (a vaccine candidate antigen) tagged with CBD were grown in transient Nicotiana benthamiana and transgenic Arabidopsis thaliana respectively. A notable fraction of the recombinant proteins was lost through plant debris pelleted from the plant-slurry made. However, this issue was resolved by adjusting tissue-to-buffer ratios with 1:10 and 1:15 in those plants respectively. Washing efficiencies were improved by agitating column beds with acidic buffer (20mM NaAc. pH 4.0) in Nicotiana and alkaline buffer (10mM Tris-base pH 8.0) in Arabidopsis. Adsorption and coupling of tagged proteins on cellulose matrices were affected by the buffer-logged resins. The column-beds, after pumping the moisture out, showed efficient in binding of antigens with almost no losses detected by immunoblot signals. The bound antigens were released efficiently from the cellulose matrices by 1% Cellobiose and 2% Triethylamine respectively. The successive purifications of these antigenic proteins with identical tags likely indicate the efficiency of the proposed strategy in providing a generic and cost-effective method to purify fusion proteins propagated in transgenic plants.

Published in	Chemical and Biomolecular Engineering (Volume 3, Issue 1)
DOI	10.11648/j.cbe.20180301.11
Page(s)	1-10
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2018. Published by Science Publishing Group

Keywords

CBD Tag, Vaccine Antigens, Affinity Separation, Plant

References

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APA Style

HMM Tariq Hossain, Kwan Yong Choi. (2018). Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants. Chemical and Biomolecular Engineering, 3(1), 1-10. https://doi.org/10.11648/j.cbe.20180301.11

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ACS Style

HMM Tariq Hossain; Kwan Yong Choi. Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants. Chem. Biomol. Eng. 2018, 3(1), 1-10. doi: 10.11648/j.cbe.20180301.11

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AMA Style

HMM Tariq Hossain, Kwan Yong Choi. Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants. Chem Biomol Eng. 2018;3(1):1-10. doi: 10.11648/j.cbe.20180301.11

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@article{10.11648/j.cbe.20180301.11,
  author = {HMM Tariq Hossain and Kwan Yong Choi},
  title = {Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants},
  journal = {Chemical and Biomolecular Engineering},
  volume = {3},
  number = {1},
  pages = {1-10},
  doi = {10.11648/j.cbe.20180301.11},
  url = {https://doi.org/10.11648/j.cbe.20180301.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.cbe.20180301.11},
  abstract = {Apart from tradition, the biospecific affinity isolation has become one of the most rapidly growing concerns with cellulose binding domains (CBDs), a high-capacity tag for cost-effective purifications of fusion proteins. We report here a new strategy optimized for isolation of two fusion proteins, the FGF-1 (a human functional protein) and the H5N1 (a vaccine candidate antigen) tagged with CBD were grown in transient Nicotiana benthamiana and transgenic Arabidopsis thaliana respectively. A notable fraction of the recombinant proteins was lost through plant debris pelleted from the plant-slurry made. However, this issue was resolved by adjusting tissue-to-buffer ratios with 1:10 and 1:15 in those plants respectively. Washing efficiencies were improved by agitating column beds with acidic buffer (20mM NaAc. pH 4.0) in Nicotiana and alkaline buffer (10mM Tris-base pH 8.0) in Arabidopsis. Adsorption and coupling of tagged proteins on cellulose matrices were affected by the buffer-logged resins. The column-beds, after pumping the moisture out, showed efficient in binding of antigens with almost no losses detected by immunoblot signals. The bound antigens were released efficiently from the cellulose matrices by 1% Cellobiose and 2% Triethylamine respectively. The successive purifications of these antigenic proteins with identical tags likely indicate the efficiency of the proposed strategy in providing a generic and cost-effective method to purify fusion proteins propagated in transgenic plants.},
 year = {2018}
}

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TY  - JOUR
T1  - Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants
AU  - HMM Tariq Hossain
AU  - Kwan Yong Choi
Y1  - 2018/01/02
PY  - 2018
N1  - https://doi.org/10.11648/j.cbe.20180301.11
DO  - 10.11648/j.cbe.20180301.11
T2  - Chemical and Biomolecular Engineering
JF  - Chemical and Biomolecular Engineering
JO  - Chemical and Biomolecular Engineering
SP  - 1
EP  - 10
PB  - Science Publishing Group
SN  - 2578-8884
UR  - https://doi.org/10.11648/j.cbe.20180301.11
AB  - Apart from tradition, the biospecific affinity isolation has become one of the most rapidly growing concerns with cellulose binding domains (CBDs), a high-capacity tag for cost-effective purifications of fusion proteins. We report here a new strategy optimized for isolation of two fusion proteins, the FGF-1 (a human functional protein) and the H5N1 (a vaccine candidate antigen) tagged with CBD were grown in transient Nicotiana benthamiana and transgenic Arabidopsis thaliana respectively. A notable fraction of the recombinant proteins was lost through plant debris pelleted from the plant-slurry made. However, this issue was resolved by adjusting tissue-to-buffer ratios with 1:10 and 1:15 in those plants respectively. Washing efficiencies were improved by agitating column beds with acidic buffer (20mM NaAc. pH 4.0) in Nicotiana and alkaline buffer (10mM Tris-base pH 8.0) in Arabidopsis. Adsorption and coupling of tagged proteins on cellulose matrices were affected by the buffer-logged resins. The column-beds, after pumping the moisture out, showed efficient in binding of antigens with almost no losses detected by immunoblot signals. The bound antigens were released efficiently from the cellulose matrices by 1% Cellobiose and 2% Triethylamine respectively. The successive purifications of these antigenic proteins with identical tags likely indicate the efficiency of the proposed strategy in providing a generic and cost-effective method to purify fusion proteins propagated in transgenic plants.
VL  - 3
IS  - 1
ER  -

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Author Information

HMM Tariq Hossain

Department of Agronomy, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh
Kwan Yong Choi

Department of Convergence Biotechnology, Pohang University of Science and Technology, Pohang, Korea

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APA Style

HMM Tariq Hossain, Kwan Yong Choi. (2018). Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants. Chemical and Biomolecular Engineering, 3(1), 1-10. https://doi.org/10.11648/j.cbe.20180301.11

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ACS Style

HMM Tariq Hossain; Kwan Yong Choi. Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants. Chem. Biomol. Eng. 2018, 3(1), 1-10. doi: 10.11648/j.cbe.20180301.11

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AMA Style

HMM Tariq Hossain, Kwan Yong Choi. Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants. Chem Biomol Eng. 2018;3(1):1-10. doi: 10.11648/j.cbe.20180301.11

Copy | Download

@article{10.11648/j.cbe.20180301.11,
  author = {HMM Tariq Hossain and Kwan Yong Choi},
  title = {Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants},
  journal = {Chemical and Biomolecular Engineering},
  volume = {3},
  number = {1},
  pages = {1-10},
  doi = {10.11648/j.cbe.20180301.11},
  url = {https://doi.org/10.11648/j.cbe.20180301.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.cbe.20180301.11},
  abstract = {Apart from tradition, the biospecific affinity isolation has become one of the most rapidly growing concerns with cellulose binding domains (CBDs), a high-capacity tag for cost-effective purifications of fusion proteins. We report here a new strategy optimized for isolation of two fusion proteins, the FGF-1 (a human functional protein) and the H5N1 (a vaccine candidate antigen) tagged with CBD were grown in transient Nicotiana benthamiana and transgenic Arabidopsis thaliana respectively. A notable fraction of the recombinant proteins was lost through plant debris pelleted from the plant-slurry made. However, this issue was resolved by adjusting tissue-to-buffer ratios with 1:10 and 1:15 in those plants respectively. Washing efficiencies were improved by agitating column beds with acidic buffer (20mM NaAc. pH 4.0) in Nicotiana and alkaline buffer (10mM Tris-base pH 8.0) in Arabidopsis. Adsorption and coupling of tagged proteins on cellulose matrices were affected by the buffer-logged resins. The column-beds, after pumping the moisture out, showed efficient in binding of antigens with almost no losses detected by immunoblot signals. The bound antigens were released efficiently from the cellulose matrices by 1% Cellobiose and 2% Triethylamine respectively. The successive purifications of these antigenic proteins with identical tags likely indicate the efficiency of the proposed strategy in providing a generic and cost-effective method to purify fusion proteins propagated in transgenic plants.},
 year = {2018}
}

Copy | Download

TY  - JOUR
T1  - Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants
AU  - HMM Tariq Hossain
AU  - Kwan Yong Choi
Y1  - 2018/01/02
PY  - 2018
N1  - https://doi.org/10.11648/j.cbe.20180301.11
DO  - 10.11648/j.cbe.20180301.11
T2  - Chemical and Biomolecular Engineering
JF  - Chemical and Biomolecular Engineering
JO  - Chemical and Biomolecular Engineering
SP  - 1
EP  - 10
PB  - Science Publishing Group
SN  - 2578-8884
UR  - https://doi.org/10.11648/j.cbe.20180301.11
AB  - Apart from tradition, the biospecific affinity isolation has become one of the most rapidly growing concerns with cellulose binding domains (CBDs), a high-capacity tag for cost-effective purifications of fusion proteins. We report here a new strategy optimized for isolation of two fusion proteins, the FGF-1 (a human functional protein) and the H5N1 (a vaccine candidate antigen) tagged with CBD were grown in transient Nicotiana benthamiana and transgenic Arabidopsis thaliana respectively. A notable fraction of the recombinant proteins was lost through plant debris pelleted from the plant-slurry made. However, this issue was resolved by adjusting tissue-to-buffer ratios with 1:10 and 1:15 in those plants respectively. Washing efficiencies were improved by agitating column beds with acidic buffer (20mM NaAc. pH 4.0) in Nicotiana and alkaline buffer (10mM Tris-base pH 8.0) in Arabidopsis. Adsorption and coupling of tagged proteins on cellulose matrices were affected by the buffer-logged resins. The column-beds, after pumping the moisture out, showed efficient in binding of antigens with almost no losses detected by immunoblot signals. The bound antigens were released efficiently from the cellulose matrices by 1% Cellobiose and 2% Triethylamine respectively. The successive purifications of these antigenic proteins with identical tags likely indicate the efficiency of the proposed strategy in providing a generic and cost-effective method to purify fusion proteins propagated in transgenic plants.
VL  - 3
IS  - 1
ER  -

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