Molecular Biotechnology

, Volume 60, Issue 12, pp 924–934 | Cite as

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

  • Yoshiki Morifuji
  • Jian XuEmail author
  • Noriko Karasaki
  • Kazuhiro Iiyama
  • Daisuke Morokuma
  • Masato Hino
  • Akitsu Masuda
  • Takumi Yano
  • Hiroaki Mon
  • Takahiro Kusakabe
  • Jae Man LeeEmail author
Original Paper


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.


Baculovirus expression vector system Serpin α1-Antitrypsin Recombinant protein Silkworm 



The NIAS-Bm-oyanagi2 cell line for propagation of recombinant BmNPVs was kindly provided by Dr. Imanishi (National Institute of Agrobiological Sciences, Japan). The MALDI-TOF MS was kindly supported by Center for Advanced Instrumental and Educational Supports (Faculty of Agriculture, Kyushu University).

Compliance with Ethical Standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

12033_2018_127_MOESM1_ESM.pdf (127 kb)
Supplemental Fig. 1 O-linked glycans analysis of rAAT. rAATs were incubated with/without O-glycosidase and analyzed by 10 % SDS-PAGE. Fetuin was used as a control. M: molecular mass markers; 1: Fetuin; 2: Fetuin incubated with O-glycosidase and Neuraminidase; 3: T3 rAAT; 4: T3 rAAT incubated with O-glycosidase; 5: T3ΔPE3P rAAT; 6: T3ΔPE3P rAAT incubated with O-glycosidase. (PDF 127 KB)


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Yoshiki Morifuji
    • 1
  • Jian Xu
    • 1
    Email author
  • Noriko Karasaki
    • 1
  • Kazuhiro Iiyama
    • 2
  • Daisuke Morokuma
    • 1
  • Masato Hino
    • 1
  • Akitsu Masuda
    • 1
  • Takumi Yano
    • 1
  • Hiroaki Mon
    • 1
  • Takahiro Kusakabe
    • 1
  • Jae Man Lee
    • 1
    Email author
  1. 1.Laboratory of Insect Genome ScienceKyushu University Graduate School of Bioresource and Bioenvironmental ScienceFukuokaJapan
  2. 2.Laboratory of Plant PathologyKyushu University Graduate School of Bioresource and Bioenvironmental ScienceFukuokaJapan

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