The Government of the United States of America, as Represented by the Secretary of Homeland Security

Max Rasmussen

Michael Puckette

John Neilan

Polynucleotide constructs that express an engineered foot-and-mouth disease (FMDV) P1 precursor protein and a non-FMDV TEV protease and methods for safe and efficient recombinant production of FMDV antigens and immunogens. Recombinant production of FMDV antigens avoids the need to culture highly-infectious FMDV, while conventional culture methods for producing FMDV antigens rely on the native FMDV 3C protease which exerts toxic effects on host cells. The inventors have developed a new system that efficiently and safely processes FMDV P1 precursor without the FMDV 3C protease, thus avoiding the toxic effects associated with use of the 3C protease. The invention is also directed to the FMDV antigens and virus-like particles produced by this system as well as to FMDV vaccines, diagnostics and other biologics.

Processing engineered FMDV P1 polypeptide using an alternative TEV protease

This application is directed generally to foot-and-mouth disease virus (FMDV) 3C proteases that have been modified by mutating a polynucleotide sequence coding for the FMDV 3C protease. The modified FMDV proteases exhibit proteolytic activity on FMDV P1 precursor protein and exhibit a reduction in one or more toxic or inhibitory properties associated with an unmodified FMDV 3C protease on a host cell used to recombinantly produce it. Vectors carrying polynucleotides encoding modified FMDV 3C protease sequences can induce production of FMDV virus-like particles in a host cell when expressed in the host cell. The modified FMDV 3C proteases can generally be used to produce immunogenic FMDV preparations capable of inducing an immune response against FMDV.

DNA vaccines against foot-and-mouth disease virus

Method for producing foot-and-mouth disease virus (FMDV) viral proteins utilizing a modified FMDV 3C protease

Vaccines and pharmaceutical compositions against foot-and-mouth disease virus

Polynucleotides encoding fusion proteins contain a secretable luciferase fused to a modified polypeptide of interest are disclosed. The polypeptide of interest has been modified to remove a native N-terminal secretion sequence and has been replaced by the secretable luciferase. One example of a modified polypeptide of interest is interferon. The polynucleotides and fusion proteins have biotherapeutic, diagnostic, and quality control applications in biotechnological, medical, and veterinary fields. Methods for producing the secretable fusion protein are also disclosed.

Fusion proteins containing luciferase and a polypeptide of interest

This application is directed generally to minicircle DNA vectors for the vaccination of foot-and-mouth disease (FMD). The transgene expression cassette in the minicircle DNA vector includes: a eukaryotic translation initiation nucleotide sequence, a mutant nucleotide sequence that encodes a foot-and-mouth disease virus (FMDV) capsid polyprotein precursor that contains at least one mutation to eliminate a restriction enzyme recognition site, a nucleotide sequence that encodes a protease that cleaves the MEW capsid polyprotein precursor into plurality of FMDV capsid proteins and a translational regulatory element to regulate the expression of the protease. The minicircle DNA vectors can be transfected directly into the cell of a mammalian host. When transfected into the mammalian host cell, virus-like particles can be produced intrinsically to stimulate the mammalian host's immune system to develop adaptive immunity toward foot-and-mouth disease.

Minicircle DNA vector vaccine platform for foot-and-mouth disease and methods thereof

This application is directed generally to minicircle DNA vectors for the vaccination of foot-and-mouth disease (FMD). The transgene expression cassette in the minicircle DNA vector includes: a eukaryotic translation initiation nucleotide sequence, a mutant nucleotide sequence that encodes a foot-and-mouth disease virus (FMDV) capsid polyprotein precursor that contains at least one mutation to eliminate a restriction enzyme recognition site, a nucleotide sequence that encodes a protease that cleaves the FMDV capsid polyprotein precursor into a plurality of FMDV capsid proteins and a translational regulatory element to regulate the expression of the protease. The minicircle DNA vectors can be transfected directly into the cell of a mammalian host. When transfected into the mammalian host cell, virus-like particles can be produced intrinsically to stimulate the mammalian host's immune system to develop adaptive immunity toward foot-and-mouth disease.

Polynucleotides encoding fusion proteins comprising interferons and luciferases which have biotherapeutic, diagnostic, and quality control applications in biotechnological, medical, and veterinary fields.

Fusion protein comprising, translation interrupter sequence, and interferon amino acid sequences

Modified foot-and-mouth disease virus 3C proteases, compositions and methods thereof

Processing of a modified foot-and-mouth disease virus P1 polypeptide by an alternative protease

Methods of making and using vaccines utilizing minicircle DNA expression vectors for production of foot-and-mouth-disease virus proteins and virus-like particles

Growing community of inventors

Guilford, CT, United States of America

Max Rasmussen