Skip to main content

Advertisement

SpringerLink
Log in

Comparison of Sepharose CL-2B and CL-4B Matrices in Purification of the Hepatitis B Surface Antigen by Affinity Chromatography

  • Full Short Communication
  • Published:
Chromatographia Aims and scope Submit manuscript

Abstract

Sepharose CL-4B and Sepharose CL-2B with immobilized CB.Hep-1 monoclonal antibody were compared for studying the matrix efficiency in the immunopurification of the recombinant hepatitis B surface antigen (rHBsAg). The elution capacities of both matrices were similar over eight chromatographic cycles, significant differences were only observed in the first purification cycles. A high percentage of the adsorbed rHBsAg was not eluted from both matrices. The rHBsAg purity was not affected by matrix characteristics (pore size and percentage of agarose) and differences between Sepharose CL-4B and Sepharose CL-2B do not provoke differences in the antibody released from the matrices under defined experimental conditions.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

References

  1. Desombere I, Willems A, Gijbels Y, Leroux-Roels G (2006) J Virol 80:3506–3514

    Article  CAS  Google Scholar 

  2. da Silva S, Alves CO, Ennes I, de Andrade S, da Silva M, da Silva ED, de Simone SG, Baroni MT (2003) J Chromatogr B 787:303–311

    Article  Google Scholar 

  3. Sánchez J, País JM, Pestana Y, López I, Masso Y, Linares M, Márquez G (2008) Biochem Eng J 38:1–8

    Article  Google Scholar 

  4. Kim WB, Kim BM, Choi EC, Kim BK (1990) J Immunol Methods 126:119–124

    Article  CAS  Google Scholar 

  5. Yarmush ML, Weiss AM, Antonsen KP, Odde DJ, Yarmush DM (1992) Biotechnol Adv 10:413–446

    Article  CAS  Google Scholar 

  6. Wilchek M, Miron T (1999) React Funct Polym 41:263–268

    Article  CAS  Google Scholar 

  7. Pharmacia Biotech. Sepharose and Sepharose CL instructions (1996) Uppsala, Sweden

  8. Hardy E, Martínez E, Diago D, Díaz R, González D, Herrera L (2000) J Biotechnol 77:157–167

    Article  CAS  Google Scholar 

  9. Reyes B, Valdés R, García C, Álvarez T, Alemán MR, Tamayo A, Padilla SA, Montero JA, Cálas G, González M, Abrahantes-Pérez MC (2006) BioPharm Int 19:50–61

    Google Scholar 

  10. Hernández R, Plana L, Gómez L, Expósito N, Valdés J, Páez R, Martínez E, Beldarraín A (2005) J Chromatogr B 816:1–6

    Article  Google Scholar 

  11. March SC, Parikh I, Cuatrecasas P (1974) Anal Biochem 60:149–152

    Article  CAS  Google Scholar 

  12. Kohn J, Wilchek M (1978) Biochem Biophys Res Commun 84:7–14

    Article  CAS  Google Scholar 

  13. Gómez L, Hernández R, Ibarra N, Valdés R, Campos Y, Tamayo A, Fernández D, Figueroa A, Álvarez T, Montero JA (2002) J Biochem Biophys Methods 52:151–159

    Article  Google Scholar 

  14. Lowry OH, Rosenbrough NJ, Farr AL, Randal RJ (1951) J Biol Chem 193:265–275

    CAS  Google Scholar 

  15. Valdés R, Reyes B, Álvarez T, García J, Montero JA, Figueroa A, Gómez L, Padilla S, Geada D, Abrahantes MC, Dorta L, Fernández D, Mendoza O, Ramírez N, Rodríguez M, Pujol M, Borroto C, Brito J (2003) Biochem Bioph Res Commun 310:742–747

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  17. Ibarra N, Caballero A, González E, Valdés R (1999) J Chromatogr B 735:271–277

    Article  CAS  Google Scholar 

  18. Wilchek M, Miron T, Kohn J (1984) Methods Enzymol 104:3–55

    Article  CAS  Google Scholar 

  19. Kohn J, Wilchek M (1982) Enzyme Microb Technol 4:161–163

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to thanks Relma Tavares de Oliveira Fernández for her inspiration and support and Rebecca Deming Ballentine for her carefully editing of the paper.

Author information

Authors and Affiliations

  1. Department of Inorganic Chemistry, Center for Engineering and Chemical Researches (CIIQ), F Street, #115 be/Ave 5th and Calzada, 10400, Havana, Cuba

    Eutimio G. Fernández

  2. Department of Monoclonal Antibody Production, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Ave 31 be/158 and 190, Playa, 10600, Havana, Cuba

    Rodolfo Valdés, Yenisley Medina, José A. Montero, Alejandro Figueroa, Williams Ferro, Yarysel Guevara, Tatiana Álvarez & Andrés Tamayo

  3. Department of Process Control, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Ave 31 be/158 and 190, Playa, 10600, Havana, Cuba

    Regla M. Somoza

  4. Department of Chemistry-Physcis, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Ave 31 be/158 and 190, Playa, 10600, Havana, Cuba

    Leonardo Canaán-Haden

  5. Direction of Quality Control, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Ave 31 be/158 and 190, Playa, 10600, Havana, Cuba

    Jorge L. López

Authors
  1. Eutimio G. Fernández
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rodolfo Valdés
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yenisley Medina
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. José A. Montero
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alejandro Figueroa
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Williams Ferro
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yarysel Guevara
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tatiana Álvarez
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Andrés Tamayo
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Regla M. Somoza
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Leonardo Canaán-Haden
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Jorge L. López
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Eutimio G. Fernández.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fernández, E.G., Valdés, R., Medina, Y. et al. Comparison of Sepharose CL-2B and CL-4B Matrices in Purification of the Hepatitis B Surface Antigen by Affinity Chromatography. Chroma 70, 1179–1183 (2009). https://doi.org/10.1365/s10337-009-1291-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1365/s10337-009-1291-9

Keywords

Search

Navigation