Fusion to elastin-like polypeptide increases production of bioactive human IFN-γ in tobacco.

Transgenic Research
Reza Heidari-JapelaghiMokhtar Jalali-Javaran

Abstract

The plant-based expression systems are now accredited as bioreactors for the high production of various biopharmaceuticals. However, low levels of agglomeration and the absence of effective procedures for purification of recombinant proteins have remained two essential obstacles in molecular farming. In this research, we have studied the production of human interferon gamma (hIFN-γ) in tobacco and analyzed the effects of elastin-like polypeptide (ELP) tag and subcellular localization on its accumulation. We report a remarkable enhancement of accumulation of the fusion proteins versus the corresponding unfused hIFN-γ proteins. Furthermore, the hIFN-γ (with and without ELP) accumulated to higher levels in the endoplasmic reticulum. The ELP fusion proteins were successfully recovered from total soluble protein with adding 2.75 M NaCl and three rounds of inverse transition cycling (ITC). The hIFN-γ was also separated from ELP with Enterokinase cleavage of the fusion protein and recovered by ITC. Inverse transition analysis indicated that the hIFN-γ-ELP variants aggregate above their inverse transition temperature and at high ionic strength. Investigation of glycosylation revealed that fused or unfused hIFN-γ proteins are N-glycosyl...Continue Reading

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Citations

Jun 3, 2021·International Journal of Molecular Sciences·Yu SawadaMotonobu Nakamura

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Datasets Mentioned

BETA
AB158612

Methods Mentioned

BETA
glycosylation
surface plasmon resonance
transgenic
electrophoresis
PCR
enzyme-linked immunosorbent assay
ELISA

Software Mentioned

SPSS
ExpOptimizer

Related Concepts

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Bioreactors (ASM)

Bioreactors are important devices or systems that utilize living cells or enzymes for a chemical process. These devices carefully control and monitor the environment factors such as pH, nutrients level, and temperature, which can affect the yield in a given process. Discover the latest research here.