Structural insight and stability of TNFR-Fc fusion protein (Etanercept) produced by using transgenic silkworms. (7th August 2020)

• Record Type: Journal Article
• Authors: Kiyoshi, Masato; Tatematsu, Ken-Ichiro; Tada, Minoru; Sezutsu, Hideki; Shibata, Hiroko; Ishii-Watabe, Akiko
• Is Part Of: Journal of biochemistry. Volume 169:Number 1(2021)
• Page Start: 25
• Page End: 33
• Publication Date: 2020-08-07
• Subjects: Fc fusion protein -- glycosylation -- physicochemical property -- silkworm -- therapeutic protein; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Electronic journals; 572.05
• DOI: 10.1093/jb/mvaa092
• Languages: English
• ISSNs: 0021-924X

Abstract

Abstract: Therapeutic proteins expressed using transgenic animals have been of great interest for several years. Especially, transgenic silkworm has been studied intensively because of its ease in handling, low-cost, high-yield and unique glycosylation patterns. However, the physicochemical property of the therapeutic protein expressed in transgenic silkworm remains elusive. Here, we constructed an expression system for the TNFR-Fc fusion protein (Etanercept) using transgenic silkworm. The TNFR-Fc fusion protein was employed to N-glycan analysis, which revealed an increased amount of afucosylated protein. Evidence from surface plasmon resonance analysis showed that the TNFR-Fc fusion protein exhibit increased binding affinity for Fcγ receptor IIIa and FcRn compared to the commercial Etanercept, emphasizing the profit of expression system using transgenic silkworm. We have further discussed the comparison of higher order structure, thermal stability and aggregation of the TNFR-Fc fusion protein. Graphical Abstract: