immunology-research-tissue-science-2018-posters

15 / 26

Journal of Transmitted Diseases and Immunity
ISSN: 2573-0320 Page 74
Volume 4
May 10-11, 2018
Frankfurt, Germany
Immunology Research 2018
Tissue Science 2018
JOINT EVENT
2 2 n d E d i t i o n o f I n t e r n a t i o n a l C o n f e r e n c e o n
Immunology and
Evolution of Infectious Diseases
&
1 2 t h E d i t i o n o f I n t e r n a t i o n a l C o n f e r e n c e o n
Tissue Engineering and
Regenerative Medicine
Insoluble and inactive protein aggregates known as inclusion
bodies are the major bottleneck in the recovery of bioactive
recombinant proteins from Escherichia coli. Strong denaturants
such as high concentrations of urea and guanidine hydrochloride
offer good solubility over a wide range of IBs but result in poor
recovery of bioactive protein. Recent studies showing the
presence of native-like secondary structure in inclusion bodies
have led to the development of mild solubilization agents
like organic solvents, alkaline pH and low concentration of
denaturants. These mild solubilization agents offer 5-6 times
better recovery of bioactive protein from inclusion bodies
than aforementioned strong denaturants. In this study, we
demonstrated the solubilisation potential of trifluoroethanol
(TFE), an organic solvent, in nine inclusion body proteins. Different
concentrations of TFE with or without low concentration of
denaturant were screened to arrive at an optimal ratio. A mixture
of 30% TFE with 3 M urea performed the best at solubilizing
maximum amounts of protein. Taking human growth hormone
(hGH) as a model protein, mode of action of TFE against strong
denaturantswas investigated using fluorescent spectroscopy and
circular dichroism. The results from these techniques suggested
the disruption of tertiary structure and stabilization of secondary
structure of protein. Furthermore, the cell number based activity
assays indicated the presence of fully functional and bioactive
protein recovered from TFE solubilized hGH inclusion bodies. We
concluded that TFE could be used as a mild solubilization agent
to recover maximum amount of bioactive protein from inclusion
body proteins.
Recovery of bioactive protein from inclusion bodies using mild
solubilisation agent
Amulya K Panda1 2, Priyank Singhvi and Vaibhav Upadhyay
1Product Development Cell (PDC) Laboratory, India
2National Institute of Immunology, India
J Transm Dis Immun 2018, Volume 2
DOI: 10.21767/2471-8084-C1-003