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Assessment of a Plantibody HB-01 Purification Strategy at Different Scales

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Abstract

Transgenic plant investigations focus on fine-regulation of the recombinant protein expression and purification strategies. In this article, preliminary experiments were done at analytical-scale to decide the best plantibody HB-01 purification strategy. Once it was assumed, the purification efficiency was assessed at different scales (10–600 kg of biomass). The plantibody purity measured by SDS-PAGE and LC-GF was over 90%, yielding 9.9 ± 6.2–18.6 ± 0.9 mg plantibody kg−1 of biomass and 39.9 ± 7.9–48.7 ± 2.1% of recovery. Significant differences were not observed among these parameters at these scales. Plant DNA contents were <3.3 ng mg−1 of plantibody, which is considered very low for the plantibody HB-01 application in the hepatitis B vaccine production.

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References

  1. Fischer R, Twyman M, Schillberg S (2003) Vaccine 21:820–825

    Article  CAS  Google Scholar 

  2. Stoger E, Sack M, Nicholson L, Fischer R, Christou P (2005) Curr Pharm Des 11:2439–2457

    Article  CAS  Google Scholar 

  3. Stoger E, Sack M, Fischer R, Christou P (2002) ). Curr Opin Biotechnol 13:161–166

    Article  CAS  Google Scholar 

  4. Kusnadi AR, Nikolov ZL, Howard JA (1997) Biotechnol Bioeng 56:473–484

    Article  CAS  Google Scholar 

  5. Meson D, Curling J (2000) BioPharm 13:48–54

    Google Scholar 

  6. Julian K-C, Pascal M, Grake MW, Christou P (2003) Nat Rev 4:794–805

    Article  Google Scholar 

  7. Gottschalk U (2005) BioPharm Int 18:1–17

    Google Scholar 

  8. Keen MJ, Hale C (1996) Cytotechnology 18:207–217

    Article  Google Scholar 

  9. Simonson L, Morton H, Reedy E (1994) Cytobios 77:159–165

    CAS  Google Scholar 

  10. Menkhaus T, Bai Y, Chenming Z, Nikolov Z, Charles G (2004) Biotechnol Prog 20:1001–1014

    Article  CAS  Google Scholar 

  11. Chen Z, Ricigliano JW, Klessin DF (1993) PNAS 90:9533–9537

    Article  CAS  Google Scholar 

  12. Desai U, Sur G, Daurnet S, Babbit R, Li Q (2002) Protein Expr Purif 25:195–202

    Article  CAS  Google Scholar 

  13. Majare M, Liluis G, Bulow L (1998) Plant Sci 134:103–114

    Article  Google Scholar 

  14. Larrick JW, Yu L, Nagtzger C, Jaiswal S, Wycoff K (2001) Biomol Eng 18:87–94

    Article  CAS  Google Scholar 

  15. Fischer R, Liao YC, Drossad J (1999) J Immunol Methods 226:1–10

    Article  CAS  Google Scholar 

  16. Ramírez N, Rodríguez M, Ayala M, Cremata J, Pérez M, Martínez A et al (2003) Biotechnol Appl Biochem 38:223–230

    Article  Google Scholar 

  17. Valdés R, Gómez L, Padilla S, Brito J, Reyes B, Alvarez T et al (2003) Biochem Biophys Res Commun 308:94–100

    Article  Google Scholar 

  18. Lowry OH, Rosenbroug NJ, Farr AL, Randal R (1951) J Biol Chem 193:265–275

    CAS  Google Scholar 

  19. Laemmli UK (1970) Nature 227:680–685

    Article  CAS  Google Scholar 

  20. Hiatt A, Cafferkey R, Bowish K (1989) Nature 342:76–78

    Article  CAS  Google Scholar 

  21. Valdés R, Reyes B, Alvarez T, García J, Montero J, Figueroa A et al (2003) Biochem Biophys Res Commun 310:742–747

    Article  Google Scholar 

  22. Milá L, García C, Padilla S, Mendoza O, Gómez L, Pujol M et al (2009) BioProcessing J 7:53–60

    Google Scholar 

  23. AAAS (2008) Science 320:473–475. ISSN 1538-8785

    Google Scholar 

  24. Balasubramaniam D, Wilkinson C, van Cott K, Zhang C (2003) J Chromatogr A 989:119–129

    Article  CAS  Google Scholar 

  25. Hiatt A, Puly M (2006) PNAS 103:14645–14646

    Article  CAS  Google Scholar 

Download references

Acknowledgments

Authors wish to thank the Process Control Department for analysis assistance, the Operation Department for installation of equipments and the Agriculture Section of Monoclonal Antibody Department for providing the vegetal biomass. Authors also thank Sarah Chinelo Igboke and Relma Tavares de Oliveira from the “Salvador Allende” Hospital, Faculty of Medicine. Havana, Cuba for manuscript editing.

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Authors and Affiliations

  1. Monoclonal Antibody Department, Center for Genetic Engineering and Biotechnology, Ave 31 be/158 and 190, Havana, 10600, Cuba

    Sigifredo Padilla, Rodolfo Valdés, Leonardo Gómez, Williams Ferro, Otto Mendoza, Cristina García, Leonardo Pasín, Yordan Issac & David Gavilán

  2. Quality Assurance Direction, Center for Genetic Engineering and Biotechnology, Ave 31 be/158 and 190, Havana, 10600, Cuba

    Lorely Milá

  3. Process Control Department, Center for Genetic Engineering and Biotechnology, Ave 31 be/158 and 190, Havana, 10600, Cuba

    Tatiana González, Raudel Sosa, Alberto Leyva & Julio Sánchez

  4. Quality Control Direction, Center for Genetic Engineering and Biotechnology, Ave 31 be/158 and 190, Havana, 10600, Cuba

    Mailín LaO & Yodelis Calvo

  5. Regulatory Affair Direction, Center for Genetic Engineering and Biotechnology, Ave 31 be/158 and 190, Havana, 10600, Cuba

    Rafael Sánchez

  6. Production Direction, Center for Genetic Engineering and Biotechnology, Ave 31 be/158 and 190, Havana, 10600, Cuba

    José Brito

  7. Tobacco Research Institute, Tumbadero Road, km 8½. San Antonio de los Baños, Havana, 3500, Cuba

    Déborah Geada

  8. Center for Engineering and Chemical Researches, F Street, # 115 be/Ave 5th and Calzada, Havana, 10600, Cuba

    Eutimio Fernández

Authors
  1. Sigifredo Padilla
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  2. Rodolfo Valdés
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  3. Leonardo Gómez
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  4. Déborah Geada
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  5. Williams Ferro
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  6. Otto Mendoza
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  7. Cristina García
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  8. Lorely Milá
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  9. Leonardo Pasín
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  10. Yordan Issac
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  11. David Gavilán
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  12. Tatiana González
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  13. Raudel Sosa
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  14. Alberto Leyva
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  15. Julio Sánchez
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  16. Mailín LaO
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  17. Yodelis Calvo
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  18. Rafael Sánchez
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  19. Eutimio Fernández
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  20. José Brito
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Corresponding author

Correspondence to Rodolfo Valdés.

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Padilla, S., Valdés, R., Gómez, L. et al. Assessment of a Plantibody HB-01 Purification Strategy at Different Scales. Chroma 70, 1673–1678 (2009). https://doi.org/10.1365/s10337-009-1388-1

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  • DOI: https://doi.org/10.1365/s10337-009-1388-1

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