Enhanced Solubilization and Purification of 3ABC
Non-structural Protein of Foot-and-Mouth Disease Virus from
BacterialInclusion Bodies
Muhammad Ashir Zia1, 2*, Muhammad
Salahuddin Shah1, 2 and Mudasser Habib1,2*
1College
of Biological Sciences, Nuclear Institute for Agriculture and
Biology college, (NIAB-C), Pakistan Institute of Engineering and
Applied Sciences, Nilore, 44000, Islamabad; 2Vaccine
Development Group, Animal Sciences Division, Nuclear Institute for
Agriculture and Biology, Jhang road, P.O box 128, Faisalabad, 38000.
Pakistan
*Corresponding author:
ashirzia68@gmail.com;
mudasserhabib@yahoo.com
Abstract
Nonstructural 3ABC protein of foot-and-mouth
disease virus (FMDV) is used to differentiate vaccinated from naturally infected
animals. The 3ABC protein is a polyprotein which is cleaved into membrane
associated 3A protein, three copies of 3B and 3Cpro mediated by virally encoded
3C protease. The expression of this protein in
E. coli results into the formation of
inclusion bodies which require solubilization in high concentration of
chaotropes and extensive refolding process prior to purification of the native
protein. Protein aggregation during refolding leads to the poor recovery of
protein in functional form. Alternatively, mild solubilization methods have been
proposed to recover the native and soluble protein from inclusion bodies present
in E. coli. In this study, 3ABC
protein was expressed predominantly as inclusion bodies using
E. coli host and solubilized in mild
non-ionic detergent followed by purification through Ni-NTA chromatography. The
protein recovery using this solubilization method, showed higher yield than
previous solubilization methods for 3ABC protein. This method also favored
higher stability of the 3ABC recombinant protein stored at different
temperatures. The reactivity of the proteins was analyzed by western blotting
and ELISA which showed their ability to use them as antigen for the development
of immunoassays. In conclusion, this study demonstrates an efficient and high
yielding purification method of this protein without refolding process than
previously described methods involving renaturation steps.
To Cite This Article:
Zia MA, Shah MS and Habib M, 2022.
Enhanced solubilization and purification
of 3ABC non-structural protein of foot-and-mouth disease virus from bacterialinclusion bodies. Pak Vet J, 42(1): 74-80.
http://dx.doi.org/10.29261/pakvetj/2021.082