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    MACHINE GUIDED DIRECTED EVOLUTION OF RAAV COMBINATORIAL CAPSID LIBRARIES
    2.
    发明公开
    MACHINE GUIDED DIRECTED EVOLUTION OF RAAV COMBINATORIAL CAPSID LIBRARIES 审中-公开

    公开(公告)号：US20230203533A1

    公开(公告)日：2023-06-29

    申请号：US17922174

    申请日：2021-04-28

    申请人： University of Florida Research Foundation, Incorporated

    发明人： Sergei Zolotukhin ,  Oleksandr Kondratov

    IPC分类号： C12N15/86 ,  C40B10/00

    CPC分类号： C12N15/86 ,  C40B10/00 ,  C12N2710/14143 ,  C12N2750/14151 ,  C12N2750/14143

    摘要： Provided herein are methods and compositions useful in the screening and production of recombinant AAV (rAAV) that have structural fitness and potency in insect cells and mammalian cells. The disclosure provides a directed evolution system for generating and selecting rAAV variants based on their Kozak sequences. In some embodiments, methods disclosed herein comprise the use of modified Kozak sequences to express AAV2 VP1 proteins in amounts that are useful for producing rAAV particles having high infectivities and/or transduction efficiencies. In some embodiments, methods of producing, and methods of screening, rAAV are provided that comprise the steps of infecting insect cells by administering rAAV to these cells, recovering rAAV-integrated DNA from these insect cells, and transfecting recovered rAAV into mammalian cells. Also provided herein are nucleic acids comprising nucleotide sequences encoding novel Kozak sequences. Further provided herein are mammalian cells, baculovirus particles and insect cells comprising these nucleic acids.

    Method of improving characteristics of proteins
    5.
    发明授权
    Method of improving characteristics of proteins 有权

    公开(公告)号：US10526597B2

    公开(公告)日：2020-01-07

    申请号：US15555276

    申请日：2016-05-16

    申请人： Full Spectrum Genetics, Inc.

    发明人： Robert DuBridge

    IPC分类号： C40B30/04 ,  C12N15/10 ,  C07K1/04 ,  C07K16/00 ,  C40B10/00 ,  C40B40/02 ,  C40B40/10

    摘要： The invention provides efficient methods for combining single-substitution libraries of nucleic acids that span and encode proteins of interest and for selecting resultant mutant proteins after expression which have improved properties or characteristics. Specifically, the methods comprising synthesizing a single substitution library for each of a plurality of domains of a protein; expressing separately each member of each single substitution library as a pre-candidate protein; selecting members of each single substitution library which encode pre-candidate proteins which exhibit an improvement in the one or more predetermined characteristics to form a selected library; shuffling members or the selected libraries in a PCR to produce a combinatorial shuffled library; expressing members of the shuffled library as candidate proteins; and selecting mutant proteins which have improved properties or characteristics.

    Methods and compositions for identifying a peptide having an intermolecular interaction with a target of interest
    6.
    发明授权
    Methods and compositions for identifying a peptide having an intermolecular interaction with a target of interest 有权

    公开(公告)号：US10160967B2

    公开(公告)日：2018-12-25

    申请号：US14923855

    申请日：2015-10-27

    申请人： AEBI LTD.

    发明人： Ilan Morad ,  Hanan Itzhaki

    IPC分类号： C12N15/10 ,  C40B10/00 ,  C40B40/02 ,  G01N33/50 ,  G01N33/569

    摘要： A system of a recombinant bacteriophage library and a target of interest complex, wherein the recombinant bacteriophage peptide library includes a plurality of peptides expressed on the surface of recombinant bacteriophages wherein each recombinant bacteriophage includes (a) a pill protein; wherein each pill protein includes (b) a peptide or polypeptide involved in an intermolecular interaction, which differs by at least one amino acid from other peptides or polypeptides in the library; and (c) a modified protease cleavage site proximal to the peptide, wherein the modified protease cleavage site is the same in each bacteriophage, the modified cleavage site having a reduced binding affinity to a protease, as compared to a non-modified cleavage site, and wherein the target of interest complex includes a protease, a flexible linker attached to the protease, and a target of interest attached to the flexible linker, wherein the target of interest participates in an intermolecular interaction.

    Nucleotides and aptamers containing boronic acid groups having biased binding to glycosylated proteins, and uses thereof
    8.
    发明授权
    Nucleotides and aptamers containing boronic acid groups having biased binding to glycosylated proteins, and uses thereof 有权
    含有与糖基化蛋白质有偏离结合的硼酸基团的核苷酸和适体及其用途

    公开(公告)号：US09096856B2

    公开(公告)日：2015-08-04

    申请号：US12669593

    申请日：2008-07-17

    申请人： Binghe Wang ,  Minyong Li ,  Zhen Huang ,  Na Lin

    发明人： Binghe Wang ,  Minyong Li ,  Zhen Huang ,  Na Lin

    IPC分类号： C12Q1/68 ,  C12N15/115 ,  C07H19/00 ,  C07H23/00 ,  C40B10/00 ,  G01N33/68 ,  C07H19/04 ,  C07H21/00 ,  C12N15/10 ,  G01N33/86

    CPC分类号： C12N15/115 ,  C07H19/04 ,  C07H21/00 ,  C07H23/00 ,  C12N15/1048 ,  C12N2310/16 ,  C12N2310/33 ,  G01N33/68 ,  G01N33/6893 ,  G01N33/86

    摘要： The present disclosure encompasses oligonucleotide aptamers selectively binding a target glycosylated polypeptide or protein, and having biased affinity for the glycan through a boronic acid linked to a nucleosidic base of a nucleotide(s). The disclosure further encompasses methods for isolating an aptamer(s) selectively binding a target glycosylated polypeptide, where, from a population of randomized oligonucleotides that have at least one nucleotide having a boronic acid label linked to a base, is selected a first subpopulation of aptamers binding to the target glycosylated polypeptide or protein. This subpopulation is then amplified without using boronic acid-modified TTP, and amplification products not binding to a target glycosylated polypeptide or protein are selected. The second subpopulation of aptamers is then amplified using boronic acid-modified TTP to provide a population of boronic acid-modified aptamers capable of selectively binding to a glycosylation site of a target polypeptide or protein. Other aspects of the disclosure encompass methods for the use of the modified aptamers to detect glycosylated species of a polypeptide or protein.

    摘要翻译： 本公开内容包括选择性结合靶糖基化多肽或蛋白质并且通过与核苷酸的核苷碱基连接的硼酸对聚糖具有偏向亲和力的寡核苷酸适体。 本公开还包括用于分离选择性结合靶糖基化多肽的适体的方法，其中从具有至少一个具有与碱基连接的硼酸标记的至少一个核苷酸的随机寡核苷酸群体中选择第一适配子亚群 与目标糖基化多肽或蛋白质结合。 然后在不使用硼酸修饰的TTP的情况下扩增该亚群，并且选择不与靶糖基化多肽或蛋白质结合的扩增产物。 然后使用硼酸修饰的TTP扩增适配体的第二亚群，以提供能够选择性结合靶多肽或蛋白质的糖基化位点的硼酸修饰的适配体群。 本公开的其它方面包括使用修饰的适体检测多肽或蛋白质的糖基化物种的方法。