Engineering foot-and-mouth disease virus serotype O IND R2/1975 for one-step purification by immobilized metal affinity chromatography
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Title |
Engineering foot-and-mouth disease virus serotype O IND R2/1975 for one-step purification by immobilized metal affinity chromatography
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Creator |
Jitendra K Biswal
Punam Bisht Saravanan Subramaniam Rajeev Ranjan Gaurav K Sharma Bramhadev Pattnaik |
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Subject |
Affinity chromatography; FMD virus; FMDV purification; His(6)-tagged FMDV; Reverse genetics.
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Description |
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.
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Date |
2021-08-01T11:55:38Z
2021-08-01T11:55:38Z 2015-09-01 |
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Type |
Journal
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Identifier |
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/51825 |
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Language |
English
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Relation |
Not Available;
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Publisher |
Elsevier
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