Multigene construct for immune-modulatory protein expression and methods of use

A technology for expressing vectors and amino acids, applied in chemical instruments and methods, biochemical equipment and methods, growth factors/growth regulators, etc., can solve the problems of insufficient production of immune regulatory proteins

CN111133109APending Publication Date: 2020-05-08ONCOSEC MEDICAL
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2020-05-08

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Abstract

Provided are expression vector constructs encoding multiple immunomodulatory proteins where each protein or component thereof can be expressed utilizing appropriate promoters and / or translation modifiers. Additional immunomodulatory proteins and genetic adjuvants containing shared tumor antigens can be added to further therapeutic potential as well as allow tracking of therapeutic treatment. Alsoprovided are methods of use for the expression vectors.
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Description

[0001] Cross References to Related Applications

[0002] This application claims U.S. Application No. 62 / 519,120, filed June 13, 2017, U.S. Application No. 62 / 582,917, filed November 7, 2017, and U.S. Application No. 62, filed February 9, 2018 / 628,917, which is hereby incorporated by reference in its entirety for all purposes.

[0003] pair via EFS References to sequence listings submitted as text files on the WEB

[0004] The sequence listing submitted electronically is also incorporated herein by reference in its entirety (filename: OM1702PCT SQ_ST25.txt; date created: June 12, 2018; file size: 45KB). technical field

[0005] Recombinant expression vectors for intratumoral delivery of three genes encoding therapeutically active multimeric and fusion polypeptides are described. Nucleic acids encoding polypeptides isolated from translational regulatory elements are provided. Methods of delivery are also provided. Background technique

[0006] E. coli plasmids have ...

Examples

Embodiment

[0129] List of examples

[0130] The subject matter disclosed herein includes but is not limited to the following examples.

[0131] 1. An expression vector comprising the nucleic acid sequence of SEQ ID NO:1.

[0132] 2. An expression vector comprising a nucleic acid encoding a polypeptide comprising amino acids at least 70% identical to the amino acid sequence of SEQ ID NO:9.

[0133] 3. The expression vector according to embodiment 2, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:9.

[0134] 4. The expression vector of embodiment 2 or 3, wherein the nucleic acid comprises a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO:8.

[0135] 5. The expression vector according to embodiment 4, wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO:8.

[0136] 6. The expression vector of embodiment 4 or 5, wherein said nucleic acid is operably linked to a nucleic acid encoding a P2A translational mod...

example

[0210] I. General approach

[0211] Standard methods in molecular biology are described. Maniatis et al., Molecular Cloning, A Laboratory Manual (1982), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Sambrook and Russell, Molecular Cloning ( 2001), 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Wu, Recombinant DNA (1993), vol. 217, Academic Press, San Diego, CA. Standard methods are also described in: Ausbel et al., "Current Protocols in Molecular Biology" (2001), Volumes 1 to 4, John Wiley and Sons, Inc. Inc.), New York, NY, which describes cloning of bacterial cells and DNA mutagenesis (vol. 1), cloning of mammalian cells and yeast (vol. 2), glycoconjugates, and protein expression (vol. 3) and Bioinformatics (Volume 4).

[0212]Methods for protein purification are described, including immunoprecipitation, chromatography, electrophoresis, centrifugation and crystallization. Coligan et al., Current Protocols in Protein Science (2000), V...