A self-adjuvanted nanoparticle based vaccine against infectious bronchitis virus

Li, Jianping; Helal, Zeinab H.; Karch, Christopher P.; Mishra, Neha; Girshick, Theodore; Garmendia, Antonio E.; Burkhard, Peter; Khan, Mazhar I.

doi:10.1371/journal.pone.0203771

Cited by 31 publications

(26 citation statements)

References 60 publications

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“…SAPNs are based on coiled-coil domains and are traditionally expressed in Escherichia coli 8 . SAPN vaccine candidates have been developed for a variety of diseases such as malaria, SARS, influenza, toxoplasmosis, and HIV-1 9,10,11,12,13,14,15,16,17,18,19 . The design of each SAPN candidate is specific to the pathogen of interest, however, the production, purification, and refolding techniques are generally broadly applicable.…”

Section: Introductionmentioning

confidence: 99%

Production of <em>E. coli</em>-expressed Self-Assembling Protein Nanoparticles for Vaccines Requiring Trimeric Epitope Presentation

Karch

Burkhard

Matyas

et al. 2019

JoVE

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Self-assembling protein nanoparticles (SAPNs) function as repetitive antigen displays and can be used to develop a wide range of vaccines for different infectious diseases. In this article we demonstrate a method to produce a SAPN core containing a six-helix bundle (SHB) assembly that is capable of presenting antigens in a trimeric conformation. We describe the expression of the SHB-SAPN in an E. coli system, as well as the necessary protein purification steps. We included an isopropanol wash step to reduce the residual bacterial lipopolysaccharide. As an indication of the protein identity and purity, the protein reacted with known monoclonal antibodies in Western blot analyses. After refolding, the size of the particles fell in the expected range (20 to 100 nm), which was confirmed by dynamic light scattering, nanoparticle tracking analysis, and transmission electron microscopy. The methodology described here is optimized for the SHB-SAPN, however, with only slight modifications it can be applied to other SAPN constructs. This method is also easily transferable to large scale production for GMP manufacturing for human vaccines. Video LinkThe video component of this article can be found at https://www.jove.com/video/60103/ 9 . This candidate was recognized by known monoclonal antibodies to HIV-1 envelope protein. Mice immunized with V1V2-SHB-SAPN raised V1V2 specific antibodies, that, most importantly, bound to gp70 V1V2, the correct conformational epitopes 9 . The SHB-SAPN core could have other functions beyond the role as a carrier for the HIV-1 V1V2 loop. Here we describe a detailed methodology for the expression, purification, refolding, and validation of the SHB-SAPN core. The sequence selection, nanoparticle design, the molecular cloning, and transformation of E. coli have been previously described 9 . Journal of Visualized Experiments August 2019 | 150 | e60103 | Page 2 of 11 Protocol 1. Expression of the SHB-SAPN Protein in E. coli BL21(DE3) 1. Mix 95 mL of component A and 5 mL of component B of the media in a 2 L sterile glass Erlenmeyer flask as per the manufacturer's instructions (see theTable of Materials). Add ampicillin to a final concentration of 100 μg/mL. 2. Inoculate the media with E. coli from a previously established glycerol stock culture. Incubate culture at 30 °C with shaking at 200 rotations per minute (rpm) for 48 h. NOTE: The used E. coli BL21 (DE3) stock contained the ampicillin resistant expression vector 23 with the SHB-SAPN gene. Although the general protocol of the media recommends 24 h of incubation at 37 °C, 48 h of incubation at 30 °C gave higher yield for SHB-SAPN. 3. Transfer the culture to two 50 mL conical tubes. Centrifuge the tubes at 4,000 x g for 10 min with a fixed angle rotor at 4 °C. Remove the supernatant and save the pellet to harvest cells. NOTE: The cell pellet can be either processed immediately or frozen at -80 °C until use. 2. Lysis of E. coli BL21(DE3) by sonication NOTE: Use nonpyrogenic plasticware and glassware baked at 250 °C for at least 30 min. Tris(2-car...

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Section: Introductionmentioning

confidence: 99%

Production of <em>E. coli</em>-expressed Self-Assembling Protein Nanoparticles for Vaccines Requiring Trimeric Epitope Presentation

Karch

Burkhard

Matyas

et al. 2019

JoVE

Self Cite

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show abstract

“…This vaccine elicits strong humoral and cellular immune response against the challenge strain, has no risk of gene integration into the host, and does not trigger an infection in the course of immunization [8]. A novel vaccine candidate based on self-assembly protein nanoparticle (SAPN) reportedly elicited high immune response, reduced tracheal virus shedding with lesser tracheal lesions [161]. IBV-Flagellin-SAPN was developed by the repeated display of the heptad repeat (HR) regions of B-cell epitopes of IBV S2 proteins in their native form.…”

Section: Immunopathology In the Hosts From Ndvmentioning

confidence: 99%

“…IBV-Flagellin-SAPN was developed by the repeated display of the heptad repeat (HR) regions of B-cell epitopes of IBV S2 proteins in their native form. The bioactive domain of flagellin was co-displayed to self-adjuvant the vaccine [161]. A self-adjuvanted vaccine peptide-based vaccine provides for improved immunogenicity with a reduced cost of vaccination.…”

Section: Immunopathology In the Hosts From Ndvmentioning

confidence: 99%

Towards Improved Use of Vaccination in the Control of Infectious Bronchitis and Newcastle Disease in Poultry: Understanding the Immunological Mechanisms

et al. 2021

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Infectious bronchitis (IB) and Newcastle disease (ND) are two important diseases of poultry and have remained a threat to the development of the poultry industry in many parts of the world. The immunology of avian has been well studied and numerous vaccines have been developed against the two viruses. Most of these vaccines are either inactivated vaccines or live attenuated vaccines. Inactivated vaccines induce weak cellular immune responses and require priming with live or other types of vaccines. Advanced technology has been used to produce several types of vaccines that can initiate prime immune responses. However, as a result of rapid genetic variations, the control of these two viral infections through vaccination has remained a challenge. Using various strategies such as combination of live attenuated and inactivated vaccines, development of IB/ND vaccines, use of DNA vaccines and transgenic plant vaccines, the problem is being surmounted. It is hoped that with increasing understanding of the immunological mechanisms in birds that are used in fighting these viruses, a more successful control of the diseases will be achieved. This will go a long way in contributing to global food security and the economic development of many developing countries, given the role of poultry in the attainment of these goals.

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“…In recent years, there has been a rise in the use of nanoparticles as vaccines against infectious pathogens [20][21][22][23][24][25][26][27] . Novel vaccine platforms such as Self-Assembling Protein Nanoparticles (SAPNs) which are composed of protein monomers that are capable of self-assembling into structures that mimic the size and shape of small viruses has been successfully developed as potent vaccine prototypes for Severe Acute Respiratory Syndrome [21] , HIV [24] , toxoplasmosis [25] , malaria [23] influenza [22,27] and infectious bronchitis virus [19] . Infectious bronchitis virus (IBV) is a highly prevalent coronavirus of chickens which is the foremost cause of economic loss to the US poultry industry affecting the performance of both meat-type and egg-laying birds.…”

Section: Use In Veterinary Vaccine Deliverymentioning

confidence: 99%

“…Protein nanoparticles are non-toxic, biodegradable, easily metabolized, and possess a good biocompatibility, greater stability, and relatively easy to prepare and easy size manipulation [1][2][3][4][5] . In biomedical sciences, protein nanoparticle is used in various targeted therapies for cancer [6,7] , tumor, cell imaging [8,9] and also as vaccine delivery system against infectious pathogens [19][20][21][22][23][24][25][26][27] . The advantage of using protein nanoparticle-based vaccine is that it prevents the premature degradation, provides good adjuvant properties, improves stability and also aids in targeted delivery of an antigen to the antigen presenting cells (APCs).…”

Section: Introductionmentioning

confidence: 99%

Fluodot Nanoparticle – A Promising Novel Delivery System For Veterinary Vaccine

2020

IJONR

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Most of the killed, inactivated, or live-attenuated pathogen vaccines are now replaced by modern vaccines containing isolated, highly purified antigenic protein subunits which are safer than live viruses or deactivated viruses. Another strategy is the development of nanoparticles which can mimic the repetitiveness, geometry, size and shape of the host-pathogen sur face and provide improved stability and long lasting immunogenicity, as well as serve as vehicles to deliver multiple copies of the antigens to the target cells. Several interesting advances have been made recently in the area of protein nanotechnology and here, we provide a concise review of one such novel nanoparticle called FluoDot which may be effectively used as a delivery system in some veterinary medicine applications.

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A self-adjuvanted nanoparticle based vaccine against infectious bronchitis virus

Cited by 31 publications

References 60 publications

Production of <em>E. coli</em>-expressed Self-Assembling Protein Nanoparticles for Vaccines Requiring Trimeric Epitope Presentation

Production of <em>E. coli</em>-expressed Self-Assembling Protein Nanoparticles for Vaccines Requiring Trimeric Epitope Presentation

Towards Improved Use of Vaccination in the Control of Infectious Bronchitis and Newcastle Disease in Poultry: Understanding the Immunological Mechanisms

Fluodot Nanoparticle – A Promising Novel Delivery System For Veterinary Vaccine

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