medigraphic.com
SPANISH

VacciMonitor

ISSN 1025-0298 (Electronic)
ISSN 1025-028X (Print)
  • Contents
  • View Archive
  • Information
    • General Information        
    • Directory
  • Publish
    • Instructions for authors        
  • medigraphic.com
    • Home
    • Journals index            
    • Register / Login
  • Mi perfil

2012, Number 2

<< Back Next >>

VacciMonitor 2012; 21 (2)

Molecular exclusion chromatography as methodology for the purification of bacteriophages

Camacho F, Puchades Y, Aguilar A, Amín N, García A, Otero O, Santiago VN
Full text How to cite this article

Language: Spanish
References: 12
Page: 6-9
PDF size: 101.50 Kb.


Key words:

Cromatography, bacteriophages, peptides, proteins.

ABSTRACT

Molecular exclusion chromatography can be used to purify filamentous phages which are the most commonly used virus in phage display technology of peptides and proteins. Bacteriophages were precipitated from culture supernatant using polyethylene glycol and they were loaded in a 100 cm high CL-4B Sepharose matrix using phosphate buffer as mobile phase. We recovered 61.3% of total phage loaded. SDS-PAGE analysis showed a high purity of these preparations. Molecular exclusion chromatography is an attractive alternative method for purifying filamentous phages.


REFERENCES

  1. Fagerlund A, Myrset AH, Kulseth MA. Construction and characterization of a 9-mer phage display pVIII-library with regulated peptide density. Appl Microbiol Biotechnol 2008;80:925–36.

  2. Santiago N, Puchades Y. Bacteriófagos: de la terapia con fagos a la biología combinatoria. Biotecnología Aplicada 2001;18(3):135-47.

  3. Uchiyama F, Tanaka Y, Minari Y, Tokui N. Designing scaffolds of peptides for phage display libraries. J Biosci Bioeng 2005;99:448-56.

  4. Perea SE, Reyes O, Puchades Y, Mendoza O, Vispo NS, Torrens I, et al. Antitumor Effect of a Novel Proapoptotic Peptide thatImpairs the Phosphorylation by the Protein Kinase 2 (Casein Kinase 2). Cancer Research 2004;64:7127-9.

  5. Folgori A, Tafi R, Meola A, Felici F, Galfre G, Cortese R, et al. A general strategy to identify mimotopes of pathological antigens using only random peptide libraries and human sera. EMBO J 1994;13:2236-43.

  6. Kehoe JW, Kay BK. Filamentous phage display in the new millennium. Chem Rev 2005;105:4056–72.

  7. Kay BK, Winter J, MacCafferty J. Phage display of peptides and proteins. A laboratory manual. San Diego: Academic Press Inc; 1996.

  8. Yamamoto RK, Alberts BM, Bezinger R, Lawhorne L, Treiber G. Rapid bacteriophage sedimentation in the presence of polyethilene glycol and its application to large scale virus purification. Virology 1970;40:734-44.

  9. Smith GP, Gingrich TR. Hydroxyapatite chromatography of phage-display virions. Biotechnique 2005;39:879-83.

  10. Zakharova MY, Kozyr AV, Ignatova AN, Vinnikov IA, Shemyakin IG, Kolesnikov AV. Purification of filamentous bacteriophage for phage display using size-exclusion chromatography. BioTechniques 2005;38:194-98.

  11. Scott JK, Smith GP. Searching for peptide ligands with an epitope library. Science 1990;249:386-90.

  12. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680-5.




2020     |     www.medigraphic.com

Mi perfil

C?MO CITAR (Vancouver)

VacciMonitor. 2012;21