Literature DB >> 24300384

Expression of functional recombinant hemagglutinin and neuraminidase proteins from the novel H7N9 influenza virus using the baculovirus expression system.

Irina Margine1, Peter Palese, Florian Krammer.   

Abstract

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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Year:  2013        PMID: 24300384      PMCID: PMC3970794          DOI: 10.3791/51112

Source DB:  PubMed          Journal:  J Vis Exp        ISSN: 1940-087X            Impact factor:   1.355


  26 in total

1.  Highly conserved protective epitopes on influenza B viruses.

Authors:  Cyrille Dreyfus; Nick S Laursen; Ted Kwaks; David Zuijdgeest; Reza Khayat; Damian C Ekiert; Jeong Hyun Lee; Zoltan Metlagel; Miriam V Bujny; Mandy Jongeneelen; Remko van der Vlugt; Mohammed Lamrani; Hans J W M Korse; Eric Geelen; Özcan Sahin; Martijn Sieuwerts; Just P J Brakenhoff; Ronald Vogels; Olive T W Li; Leo L M Poon; Malik Peiris; Wouter Koudstaal; Andrew B Ward; Ian A Wilson; Jaap Goudsmit; Robert H E Friesen
Journal:  Science       Date:  2012-08-09       Impact factor: 47.728

2.  Chimeric hemagglutinin influenza virus vaccine constructs elicit broadly protective stalk-specific antibodies.

Authors:  Florian Krammer; Natalie Pica; Rong Hai; Irina Margine; Peter Palese
Journal:  J Virol       Date:  2013-04-10       Impact factor: 5.103

3.  H3N2 influenza virus infection induces broadly reactive hemagglutinin stalk antibodies in humans and mice.

Authors:  Irina Margine; Rong Hai; Randy A Albrecht; Gerlinde Obermoser; A Carson Harrod; Jacques Banchereau; Karolina Palucka; Adolfo García-Sastre; Peter Palese; John J Treanor; Florian Krammer
Journal:  J Virol       Date:  2013-02-13       Impact factor: 5.103

4.  B cell response and hemagglutinin stalk-reactive antibody production in different age cohorts following 2009 H1N1 influenza virus vaccination.

Authors:  Mark Y Sangster; Jane Baer; Felix W Santiago; Theresa Fitzgerald; Natalia A Ilyushina; Aarthi Sundararajan; Alicia D Henn; Florian Krammer; Hongmei Yang; Catherine J Luke; Martin S Zand; Peter F Wright; John J Treanor; David J Topham; Kanta Subbarao
Journal:  Clin Vaccine Immunol       Date:  2013-04-10

5.  Human infection with a novel avian-origin influenza A (H7N9) virus.

Authors:  Rongbao Gao; Bin Cao; Yunwen Hu; Zijian Feng; Dayan Wang; Wanfu Hu; Jian Chen; Zhijun Jie; Haibo Qiu; Ke Xu; Xuewei Xu; Hongzhou Lu; Wenfei Zhu; Zhancheng Gao; Nijuan Xiang; Yinzhong Shen; Zebao He; Yong Gu; Zhiyong Zhang; Yi Yang; Xiang Zhao; Lei Zhou; Xiaodan Li; Shumei Zou; Ye Zhang; Xiyan Li; Lei Yang; Junfeng Guo; Jie Dong; Qun Li; Libo Dong; Yun Zhu; Tian Bai; Shiwen Wang; Pei Hao; Weizhong Yang; Yanping Zhang; Jun Han; Hongjie Yu; Dexin Li; George F Gao; Guizhen Wu; Yu Wang; Zhenghong Yuan; Yuelong Shu
Journal:  N Engl J Med       Date:  2013-04-11       Impact factor: 91.245

6.  Hemagglutinin stalk antibodies elicited by the 2009 pandemic influenza virus as a mechanism for the extinction of seasonal H1N1 viruses.

Authors:  Natalie Pica; Rong Hai; Florian Krammer; Taia T Wang; Jad Maamary; Dirk Eggink; Gene S Tan; Jens C Krause; Thomas Moran; Cheryl R Stein; David Banach; Jens Wrammert; Robert B Belshe; Adolfo García-Sastre; Peter Palese
Journal:  Proc Natl Acad Sci U S A       Date:  2012-01-30       Impact factor: 11.205

7.  A pan-H1 anti-hemagglutinin monoclonal antibody with potent broad-spectrum efficacy in vivo.

Authors:  Gene S Tan; Florian Krammer; Dirk Eggink; Alita Kongchanagul; Thomas M Moran; Peter Palese
Journal:  J Virol       Date:  2012-04-04       Impact factor: 5.103

8.  1976 and 2009 H1N1 influenza virus vaccines boost anti-hemagglutinin stalk antibodies in humans.

Authors:  Matthew S Miller; Tshidi Tsibane; Florian Krammer; Rong Hai; Saad Rahmat; Christopher F Basler; Peter Palese
Journal:  J Infect Dis       Date:  2012-10-18       Impact factor: 5.226

9.  A carboxy-terminal trimerization domain stabilizes conformational epitopes on the stalk domain of soluble recombinant hemagglutinin substrates.

Authors:  Florian Krammer; Irina Margine; Gene S Tan; Natalie Pica; Jens C Krause; Peter Palese
Journal:  PLoS One       Date:  2012-08-23       Impact factor: 3.240

10.  Influenza virus infection in guinea pigs raised as livestock, Ecuador.

Authors:  Victor H Leyva-Grado; Samira Mubareka; Florian Krammer; Washington B Cárdenas; Peter Palese
Journal:  Emerg Infect Dis       Date:  2012-07       Impact factor: 6.883

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  94 in total

1.  A Multi-Targeting, Nucleoside-Modified mRNA Influenza Virus Vaccine Provides Broad Protection in Mice.

Authors:  Alec W Freyn; Jamile Ramos da Silva; Victoria C Rosado; Carly M Bliss; Matthew Pine; Barbara L Mui; Ying K Tam; Thomas D Madden; Luís Carlos de Souza Ferreira; Drew Weissman; Florian Krammer; Lynda Coughlan; Peter Palese; Norbert Pardi; Raffael Nachbagauer
Journal:  Mol Ther       Date:  2020-04-19       Impact factor: 11.454

2.  Broadly Cross-Reactive, Nonneutralizing Antibodies against Influenza B Virus Hemagglutinin Demonstrate Effector Function-Dependent Protection against Lethal Viral Challenge in Mice.

Authors:  Guha Asthagiri Arunkumar; Andriani Ioannou; Teddy John Wohlbold; Philip Meade; Sadaf Aslam; Fatima Amanat; Juan Ayllon; Adolfo García-Sastre; Florian Krammer
Journal:  J Virol       Date:  2019-03-05       Impact factor: 5.103

3.  Assessment of influenza virus hemagglutinin stalk-based immunity in ferrets.

Authors:  Florian Krammer; Rong Hai; Mark Yondola; Gene S Tan; Victor H Leyva-Grado; Alex B Ryder; Matthew S Miller; John K Rose; Peter Palese; Adolfo García-Sastre; Randy A Albrecht
Journal:  J Virol       Date:  2014-01-08       Impact factor: 5.103

4.  Monoclonal Antibody Therapy Protects Pharmacologically Immunosuppressed Mice from Lethal Infection with Influenza B Virus.

Authors:  Bindumadhav M Marathe; Guha Asthagiri Arunkumar; Peter Vogel; Philippe Noriel Q Pascua; Jeremy Jones; Richard J Webby; Florian Krammer; Elena A Govorkova
Journal:  Antimicrob Agents Chemother       Date:  2020-08-20       Impact factor: 5.191

5.  Hemagglutinin Stalk- and Neuraminidase-Specific Monoclonal Antibodies Protect against Lethal H10N8 Influenza Virus Infection in Mice.

Authors:  Teddy John Wohlbold; Veronika Chromikova; Gene S Tan; Philip Meade; Fatima Amanat; Phillip Comella; Ariana Hirsh; Florian Krammer
Journal:  J Virol       Date:  2015-10-28       Impact factor: 5.103

6.  Development of Influenza B Universal Vaccine Candidates Using the "Mosaic" Hemagglutinin Approach.

Authors:  Weina Sun; Ericka Kirkpatrick; Megan Ermler; Raffael Nachbagauer; Felix Broecker; Florian Krammer; Peter Palese
Journal:  J Virol       Date:  2019-05-29       Impact factor: 5.103

7.  Preexisting human antibodies neutralize recently emerged H7N9 influenza strains.

Authors:  Carole J Henry Dunand; Paul E Leon; Kaval Kaur; Gene S Tan; Nai-Ying Zheng; Sarah Andrews; Min Huang; Xinyan Qu; Yunping Huang; Marlene Salgado-Ferrer; Irvin Y Ho; William Taylor; Rong Hai; Jens Wrammert; Rafi Ahmed; Adolfo García-Sastre; Peter Palese; Florian Krammer; Patrick C Wilson
Journal:  J Clin Invest       Date:  2015-02-17       Impact factor: 14.808

8.  Microcrystallography of Protein Crystals and In Cellulo Diffraction.

Authors:  Marion Boudes; Damià Garriga; Fasséli Coulibaly
Journal:  J Vis Exp       Date:  2017-07-21       Impact factor: 1.355

9.  Newcastle Disease Virus-Vectored H7 and H5 Live Vaccines Protect Chickens from Challenge with H7N9 or H5N1 Avian Influenza Viruses.

Authors:  Qinfang Liu; Ignacio Mena; Jingjiao Ma; Bhupinder Bawa; Florian Krammer; Young S Lyoo; Yuekun Lang; Igor Morozov; Gusti Ngurah Mahardika; Wenjun Ma; Adolfo García-Sastre; Juergen A Richt
Journal:  J Virol       Date:  2015-04-29       Impact factor: 5.103

10.  Extending the Stalk Enhances Immunogenicity of the Influenza Virus Neuraminidase.

Authors:  Felix Broecker; Allen Zheng; Nungruthai Suntronwong; Weina Sun; Mark J Bailey; Florian Krammer; Peter Palese
Journal:  J Virol       Date:  2019-08-28       Impact factor: 5.103

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