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    GEMINIVIRAL VECTORS THAT REDUCE CELL DEATH AND ENHANCE EXPRESSION OF BIOPHARMACEUTICAL PROTEINS
    4.
    发明申请
    GEMINIVIRAL VECTORS THAT REDUCE CELL DEATH AND ENHANCE EXPRESSION OF BIOPHARMACEUTICAL PROTEINS 有权

    公开(公告)号：US20220235362A1

    公开(公告)日：2022-07-28

    申请号：US17607843

    申请日：2020-04-30

    Applicant: Arizona Board of Regents on behalf of Arizona State University

    Inventor： Hugh Mason ,  Andrew Diamos

    IPC: C12N15/82 ,  C12N1/20 ,  C12N15/74

    Abstract: The disclosure relates to a T-DNA binary vector based on bean yellow dwarf virus (BeYDV) that reduces plant cell death and increases transgene expression. In one aspect, the T-DNA region comprise a replicon cassette comprising a rep gene or a repA gene with a mutated translation initiation region. The disclosure also relates to replicating geminiviral expression system based on BeYDV comprising with an expression cassette a sequence encoding Rep and a sequence encoding the promoter of ubiquitin-3 from potato with ubiquitin fusion; an expression cassette comprising a sequence encoding RepA and a sequence encoding the promoter of ubiquitin-3 from potato with ubiquitin fusion; and an expression cassette comprising a promoter region, a 5′ UTR, a sequence encoding a recombinant protein, and a 3′ UTR. These expression cassettes are on different T-DNA cloning vectors or on one T-DNA cloning vector.

    OPTIMIZED PLANT EXPRESSION SYSTEMS FOR HIGH LEVEL PRODUCTION OF MONOCLONAL ANTIBODIES AND METHODS OF PRODUCTION THEREOF
    5.
    发明申请
    OPTIMIZED PLANT EXPRESSION SYSTEMS FOR HIGH LEVEL PRODUCTION OF MONOCLONAL ANTIBODIES AND METHODS OF PRODUCTION THEREOF 有权

    公开(公告)号：US20210198693A1

    公开(公告)日：2021-07-01

    申请号：US17190745

    申请日：2021-03-03

    Applicant: ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY

    Inventor： Hugh Mason ,  Mary Pardhe ,  Andrew Diamos

    IPC: C12N15/86 ,  C12N15/82

    Abstract: Improved plant transient expression systems using optimized geminiviral vectors that efficiently produce heteromultimeric proteins are described herein. Examples of high yields are shown herein, including two, three, or four fluorescent proteins coexpressed simultaneously. Various antibodies were produced using the optimized vectors with special focus given to the creation and production of a chimeric broadly neutralizing anti-flavivirus antibody. The variable regions of this murine antibody, 2A10G6, were codon optimized and fused to a human IgG1. Analysis of the chimeric antibody showed that it was efficiently expressed in plants, can be purified to near homogeneity by a simple one-step purification process, retains its ability to recognize the Zika virus envelope protein, and induce an immune response against Zika virus. Two other monoclonal antibodies were produced at similar levels. This technology is versatile tool for the production of a wide spectrum of pharmaceutical multi-protein complexes in a fast, powerful, and cost-effective way.

    Optimized plant expression systems
    6.
    发明授权
    Optimized plant expression systems 有权

    公开(公告)号：US12297443B2

    公开(公告)日：2025-05-13

    申请号：US17190745

    申请日：2021-03-03

    Applicant: ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY

    Inventor： Hugh Mason ,  Mary Pardhe ,  Andrew Diamos

    IPC: C12N15/86 ,  C12N15/82

    Abstract: Improved plant transient expression systems using optimized geminiviral vectors that efficiently produce heteromultimeric proteins are described herein. Examples of high yields are shown herein, including two, three, or four fluorescent proteins coexpressed simultaneously. Various antibodies were produced using the optimized vectors with special focus given to the creation and production of a chimeric broadly neutralizing anti-flavivirus antibody. The variable regions of this murine antibody, 2A10G6, were codon optimized and fused to a human IgG1. Analysis of the chimeric antibody showed that it was efficiently expressed in plants, can be purified to near homogeneity by a simple one-step purification process, retains its ability to recognize the Zika virus envelope protein, and induce an immune response against Zika virus. Two other monoclonal antibodies were produced at similar levels. This technology is versatile tool for the production of a wide spectrum of pharmaceutical multi-protein complexes in a fast, powerful, and cost-effective way.

    Replicating and non-replicating vectors for recombinant protein production in plants and method of use thereof
    10.
    发明授权
    Replicating and non-replicating vectors for recombinant protein production in plants and method of use thereof 有权

    公开(公告)号：US12286636B2

    公开(公告)日：2025-04-29

    申请号：US16976739

    申请日：2019-03-04

    Applicant: ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY

    Inventor： Hugh Mason ,  Andrew Diamos ,  Sun Hee Rosenthal

    IPC: C12N15/82

    Abstract: The present disclosure relates to plant-based recombinant protein production systems and their methods of production and use. The plant-based recombinant protein production system is a vector comprising a 5′ UTR and a 3′ UTR, wherein the 3′ UTR comprises at least one terminator selected from the group consisting of: EU, IEU, NbACT3, NbACT617, NbACT567, Pin2, BDB501, BDB282, NbHSP, NbHSPb, Rep, RbcS, SIR, SIR 5′/3′, SIR 3′, AtHSP, 35S, RepA, NOS, TMV, TNVD, PEMV, and BYDV. In certain implementations, the vector comprises two terminators in the 3′ UTR, where the two terminators are fused to form a double terminator. For example, the double terminator comprises two members selected from the group consisting of: EU, IEU, NbACT3, NbACT617, NbACT567, Pin2, BDB501, BDB282, NbHSP, NbHSPb, Rep, RbcS, SIR, SIR 5′/3′, SIR 3′, AtHSP, 35S, RepA, NOS, TMV, TNVD, PEMV, and BYDV. In some aspects, the vector further comprises a chromatin scaffold/matrix attachment region (MAR) that is downstream of the terminators.

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