Engineering foot-and-mouth disease virus serotype O IND R2/1975 for one-step purification by immobilized metal affinity chromatography

Jitendra K. Biswal, Punam Bisht, Saravanan Subramaniam, Rajeev Ranjan, Gaurav K. Sharma, Bramhadev Pattnaik

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Immobilized metal affinity chromatography (IMAC) allows for the efficient protein purification via metal affinity tag such as hexa-histidine (His6) sequence. To develop a new chromatography strategy for the purification and concentration of foot-and-mouth disease virus (FMDV) particles, we inserted the His6-tag at the earlier reported site in the VP1 G-H loop of the FMD virus serotype O vaccine strain IND R2/1975. Display of the His6-tag on the capsid surface, endowed the virus with an increased affinity for immobilized nickel ions. We demonstrated that the His6-tagged FMDV could be produced to high titre and purified from the infected BHK-21 cell lysates by IMAC efficiently. Further, a 1150-fold reduction in protein contaminant level and an 8400-fold reduction in DNA contaminant level were achieved in the IMAC purification of His6-tagged FMDV. Through various functional assays it has been found that the tagged virus retains its functionality and infectivity similar to the non-tagged virus. The affinity purification of the His6-tagged FMDV may offer a feasible, alternative approach to the current methods of FMDV antigen purification, concentration and process scalability.

Original languageEnglish
Pages (from-to)390-398
Number of pages9
JournalBiologicals
Volume43
Issue number5
DOIs
StatePublished - 1 Sep 2015
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2015 The International Alliance for Biological Standardization.

Keywords

  • Affinity chromatography
  • FMD virus
  • FMDV purification
  • His-tagged FMDV
  • Reverse genetics

Fingerprint

Dive into the research topics of 'Engineering foot-and-mouth disease virus serotype O IND R2/1975 for one-step purification by immobilized metal affinity chromatography'. Together they form a unique fingerprint.

Cite this