Expression of Functional Recombinant Hemagglutinin and Neuraminidase Proteins from the Novel H7N9 Influenza Virus Using the Baculovirus Expression System

被引:150
作者
Margine, Irina [1 ,2 ]
Palese, Peter [1 ,3 ]
Krammer, Florian [1 ]
机构
[1] Icahn Sch Med Mt Sinai, Dept Microbiol, New York, NY 10029 USA
[2] Icahn Sch Med Mt Sinai, Grad Sch Biol Sci, New York, NY 10029 USA
[3] Icahn Sch Med Mt Sinai, Dept Med, New York, NY 10029 USA
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2013年 / 81期
基金
奥地利科学基金会;
关键词
Infection; Issue; 81; Influenza A virus; Orthomyxoviridae Infections; Influenza; Human; Influenza in Birds; Influenza Vaccines; hemagglutinin; neuraminidase; H7N9; baculovirus; insect cells; recombinant protein expression;
D O I
10.3791/51112
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The baculovirus expression system is a powerful tool for expression of recombinant proteins. Here we use it to produce correctly folded and glycosylated versions of the influenza A virus surface glycoproteins - the hemagglutinin (HA) and the neuraminidase (NA). As an example, we chose the HA and NA proteins expressed by the novel H7N9 virus that recently emerged in China. However the protocol can be easily adapted for HA and NA proteins expressed by any other influenza A and B virus strains. Recombinant HA (rHA) and NA (rNA) proteins are important reagents for immunological assays such as ELISPOT and ELISA, and are also in wide use for vaccine standardization, antibody discovery, isolation and characterization. Furthermore, recombinant NA molecules can be used to screen for small molecule inhibitors and are useful for characterization of the enzymatic function of the NA, as well as its sensitivity to antivirals. Recombinant HA proteins are also being tested as experimental vaccines in animal models, and a vaccine based on recombinant HA was recently licensed by the FDA for use in humans. The method we describe here to produce these molecules is straight forward and can facilitate research in influenza laboratories, since it allows for production of large amounts of proteins fast and at a low cost. Although here we focus on influenza virus surface glycoproteins, this method can also be used to produce other viral and cellular surface proteins.
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页数:10
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共 26 条
[1]   Development of recombinant protein-based influenza vaccine Expression and affinity purification of H1N1 influenza virus neuraminidase [J].
Dalakouras, Thanasis ;
Smith, Brian J. ;
Platis, Dimitris ;
Cox, Manon M. J. ;
Labrou, Nikolaos E. .
JOURNAL OF CHROMATOGRAPHY A, 2006, 1136 (01) :48-56
[2]   Highly Conserved Protective Epitopes on Influenza B Viruses [J].
Dreyfus, Cyrille ;
Laursen, Nick S. ;
Kwaks, Ted ;
Zuijdgeest, David ;
Khayat, Reza ;
Ekiert, Damian C. ;
Lee, Jeong Hyun ;
Metlagel, Zoltan ;
Bujny, Miriam V. ;
Jongeneelen, Mandy ;
van der Vlugt, Remko ;
Lamrani, Mohammed ;
Korse, Hans J. W. M. ;
Geelen, Eric ;
Sahin, Ozcan ;
Sieuwerts, Martijn ;
Brakenhoff, Just P. J. ;
Vogels, Ronald ;
Li, Olive T. W. ;
Poon, Leo L. M. ;
Peiris, Malik ;
Koudstaal, Wouter ;
Ward, Andrew B. ;
Wilson, Ian A. ;
Goudsmit, Jaap ;
Friesen, Robert H. E. .
SCIENCE, 2012, 337 (6100) :1343-1348
[3]   Antibody Recognition of a Highly Conserved Influenza Virus Epitope [J].
Ekiert, Damian C. ;
Bhabha, Gira ;
Elsliger, Marc-Andre ;
Friesen, Robert H. E. ;
Jongeneelen, Mandy ;
Throsby, Mark ;
Goudsmit, Jaap ;
Wilson, Ian A. .
SCIENCE, 2009, 324 (5924) :246-251
[4]   Avian influenza A virus (H7N7) associated with human conjunctivitis and a fatal case of acute respiratory distress syndrome. [J].
Fouchier, RAM ;
Schneeberger, PM ;
Rozendaal, FW ;
Broekman, JM ;
Kemink, SAG ;
Munstert, V ;
Kuiken, T ;
Rimmelzwaan, GF ;
Schutten, M ;
van Doornum, GJJ ;
Koch, G ;
Bosman, A ;
Koopmans, M ;
Osterhaus, ADME .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (05) :1356-1361
[5]   Human Infection with a Novel Avian-Origin Influenza A (H7N9) Virus [J].
Gao, Rongbao ;
Cao, Bin ;
Hu, Yunwen ;
Feng, Zijian ;
Wang, Dayan ;
Hu, Wanfu ;
Chen, Jian ;
Jie, Zhijun ;
Qiu, Haibo ;
Xu, Ke ;
Xu, Xuewei ;
Lu, Hongzhou ;
Zhu, Wenfei ;
Gao, Zhancheng ;
Xiang, Nijuan ;
Shen, Yinzhong ;
He, Zebao ;
Gu, Yong ;
Zhang, Zhiyong ;
Yang, Yi ;
Zhao, Xiang ;
Zhou, Lei ;
Li, Xiaodan ;
Zou, Shumei ;
Zhang, Ye ;
Li, Xiyan ;
Yang, Lei ;
Guo, Junfeng ;
Dong, Jie ;
Li, Qun ;
Dong, Libo ;
Zhu, Yun ;
Bai, Tian ;
Wang, Shiwen ;
Hao, Pei ;
Yang, Weizhong ;
Zhang, Yanping ;
Han, Jun ;
Yu, Hongjie ;
Li, Dexin ;
Gao, George F. ;
Wu, Guizhen ;
Wang, Yu ;
Yuan, Zhenghong ;
Shu, Yuelong .
NEW ENGLAND JOURNAL OF MEDICINE, 2013, 368 (20) :1888-1897
[6]   Induction of Cross-Reactive Antibodies to Novel H7N9 Influenza Virus by Recombinant Newcastle Disease Virus Expressing a North American Lineage H7 Subtype Hemagglutinin [J].
Goff, Peter H. ;
Krammer, Florian ;
Hai, Rong ;
Seibert, Christopher W. ;
Margine, Irina ;
Garcia-Sastre, Adolfo ;
Palese, Peter .
JOURNAL OF VIROLOGY, 2013, 87 (14) :8235-8240
[7]   Baculovirus as versatile vectors for protein expression in insect and mammalian cells [J].
Kost, TA ;
Condreay, JP ;
Jarvis, DL .
NATURE BIOTECHNOLOGY, 2005, 23 (05) :567-575
[8]  
Krammer F, 2013, J VIROL, V87, P6542, DOI [10.1128/JVI.03183-13, 10.1128/JVI.00641-13]
[9]   A Carboxy-Terminal Trimerization Domain Stabilizes Conformational Epitopes on the Stalk Domain of Soluble Recombinant Hemagglutinin Substrates [J].
Krammer, Florian ;
Margine, Irina ;
Tan, Gene S. ;
Pica, Natalie ;
Krause, Jens C. ;
Palese, Peter .
PLOS ONE, 2012, 7 (08)
[10]   Alternative influenza vaccines made by insect cells [J].
Krammer, Florian ;
Grabherr, Reingard .
TRENDS IN MOLECULAR MEDICINE, 2010, 16 (07) :313-320