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Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines
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Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines
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Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines
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Journal Article
Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines
2014
Overview
The use of DNA to deliver vaccine antigens offers many advantages, including ease of manufacture and cost. However, most DNA vaccines are plasmids and must be grown in bacterial culture, necessitating elements that are either unnecessary for effective gene delivery (for example, bacterial origins of replication) or undesirable (for example, antibiotic resistance genes). Removing these elements may improve the safety profile of DNA for the delivery of vaccines. Here, we describe a novel, double-stranded, linear DNA construct produced by an enzymatic process that solely encodes an antigen expression cassette, comprising antigen, promoter, polyA tail and telomeric ends. We compared these constructs (called ‘Doggybones’ because of their shape) with conventional plasmid DNA. Using luciferase-expressing constructs, we demonstrated that expression levels were equivalent between Doggybones and plasmids both
in vitro
and
in vivo
. When mice were immunized with DNA constructs expressing the HIV envelope protein gp140, equivalent humoral and cellular responses were induced. Immunizations with either construct type expressing hemagluttinin were protective against H1N1 influenza challenge. This is the first example of an effective DNA vaccine, which can be produced on a large scale by enzymatic processes.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ 38/5
/ 59/5
/ 64/60
/ Animals
/ Antigens
/ Biomedical and Life Sciences
/ DNA
/ Dogs
/ env Gene Products, Human Immunodeficiency Virus - genetics
/ env Gene Products, Human Immunodeficiency Virus - immunology
/ Enzymes
/ Female
/ Genetic Vectors - administration & dosage
/ Genetic Vectors - immunology
/ Hemagglutinins, Viral - genetics
/ Hemagglutinins, Viral - immunology
/ HIV
/ Human immunodeficiency virus
/ Influenza A Virus, H1N1 Subtype - immunology
/ Influenza A Virus, H1N1 Subtype - physiology
/ Madin Darby Canine Kidney Cells
/ Methods
/ Mice
/ Orthomyxoviridae Infections - immunology
/ Orthomyxoviridae Infections - virology
/ Plasmids
/ Testing
/ Vaccines
