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Expression, Purification, and Characterization of Recombinant Human α1-Antitrypsin Produced Using Silkworm–Baculovirus Expression System

Author:
Morifuji, Yoshiki, Xu, Jian, Karasaki, Noriko, Iiyama, Kazuhiro, Morokuma, Daisuke, Hino, Masato, Masuda, Akitsu, Yano, Takumi, Mon, Hiroaki, Kusakabe, Takahiro, Lee, Jae Man
Source:
Molecular biotechnology 2018 v.60 no.12 pp. 924-934
ISSN:
1073-6085
Subject:
Baculoviridae, blood serum, enzyme activity, genetic vectors, humans, insect larvae, serine proteinase inhibitors, silkworms
Abstract:
Human α₁-antitrypsin (AAT) is the most abundant serine proteinase inhibitor (serpin) in the human plasma. Commercially available AAT for the medications of deficiency of α₁-antitrypsin is mainly purified from human plasma. There is a high demand for a stable and low-cost supply of recombinant AAT (rAAT). In this study, the baculovirus expression vector system using silkworm larvae as host was employed and a large amount of highly active AAT was recovered from the silkworm serum (~ 15 mg/10 ml) with high purity. Both the enzymatic activity and stability of purified rAAT were comparable with those of commercial product. Our results provide an alternative method for mass production of the active rAAT in pharmaceutical use.
Agid:
6195253