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 language | English |
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Pages (from-to) | 390-398 |
Number of pages | 9 |
Journal | Biologicals |
Volume | 43 |
Issue number | 5 |
DOIs | |
State | Published - 1 Sep 2015 |
Externally published | Yes |
Bibliographical note
Publisher Copyright:© 2015 The International Alliance for Biological Standardization.
Funding
This work was supported by Indian Council of Agricultural Research under the project number-IXX08486.
Funders | Funder number |
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Indian Council of Agricultural Research |
Keywords
- Affinity chromatography
- FMD virus
- FMDV purification
- His-tagged FMDV
- Reverse genetics