INTRINSICALLY DISORDERED PROTEINS FOR EXTRACTION OF NUCLEIC ACIDS

    公开(公告)号:US20230407367A1

    公开(公告)日:2023-12-21

    申请号:US18137921

    申请日:2023-04-21

    IPC分类号: C12Q1/6806

    CPC分类号: C12Q1/6806

    摘要: The present disclosure relates to compositions and methods for isolating nucleic acids from a sample comprising nucleic acids, such as physiologically relevant samples or environmental samples. The sample can be incubated with a population of polymers that bind the nucleic acids to form a coacervate in a liquid-liquid phase separated solution. Incubation can be performed at a temperature above a concentration dependent phase separation transition temperature of the polymers. The resulting coacervate can be decanted from the liquid-liquid phase separated solution. The nucleic acids can be separated from the polymers by adding a salt solution, adjusting the pH, or both to disrupt an electrostatic interaction between the nucleic acids and the polymers.

    Affinity selection of Nipah and Hendra virus-related vaccine candidates from a complex random peptide library displayed on bacteriophage virus-like particles
    2.
    发明授权
    Affinity selection of Nipah and Hendra virus-related vaccine candidates from a complex random peptide library displayed on bacteriophage virus-like particles 有权
    从噬菌体病毒样颗粒中显示的复杂随机肽库的Nipah和Hendra病毒相关疫苗候选物的亲和力选择

    公开(公告)号:US09549976B1

    公开(公告)日:2017-01-24

    申请号:US14081629

    申请日:2013-11-15

    摘要: The invention relates to virus-like particles of bacteriophage MS2 (MS2 VLPs) displaying peptide epitopes or peptide mimics of epitopes of Nipah Virus envelope glycoprotein that elicit an immune response against Nipah Virus upon vaccination of humans or animals. Affinity selection on Nipah Virus-neutralizing monoclonal antibodies using random sequence peptide libraries on MS2 VLPs selected peptides with sequence similarity to peptide sequences found within the envelope glycoprotein of Nipah itself, thus identifying the epitopes the antibodies recognize. The selected peptide sequences themselves are not necessarily identical in all respects to a sequence within Nipah Virus glycoprotein, and therefore may be referred to as epitope mimics VLPs displaying these epitope mimics can serve as vaccine. On the other hand, display of the corresponding wild-type sequence derived from Nipah Virus and corresponding to the epitope mapped by affinity selection, may also be used as a vaccine.

    摘要翻译: 本发明涉及噬菌体MS2(MS2 VLP)的病毒样颗粒,其显示了尼泊尔病毒包膜糖蛋白的表位的肽表位或肽模拟物,其在接种人或动物时引起针对尼帕病毒的免疫应答。 在Nipah病毒中和单克隆抗体上的亲和力选择,使用MS2 VLP上的随机序列肽文库,选择与Nipah本身的包膜糖蛋白内发现的肽序列具有序列相似性的肽,从而鉴定抗体识别的表位。 所选择的肽序列本身在所有方面不一定与尼帕病毒糖蛋白内的序列相同,因此可以称为表位模拟物,显示这些表位模拟物的VLP可以用作疫苗。 另一方面,从Nipah Virus得到的相应野生型序列的显示与通过亲和力选择映射的表位对应,也可以用作疫苗。

    Methods for Screening Viral Like Particles and Identifying Neutralizing Epitopes and Related Vaccines, Constructs, and Libraries
    4.
    发明申请
    Methods for Screening Viral Like Particles and Identifying Neutralizing Epitopes and Related Vaccines, Constructs, and Libraries 审中-公开
    筛选病毒颗粒和鉴定中和表位和相关疫苗,构建体和图书馆的方法

    公开(公告)号:US20130149336A1

    公开(公告)日:2013-06-13

    申请号:US13662060

    申请日:2012-10-26

    IPC分类号: G01N33/68

    摘要: The invention is directed to methods of screening immunogenic viral like particles and related immunogenic compositions and diagnostic techniques. In one embodiment, the invention provides methods of screening immunogenic viral like particles containing peptides corresponding to epitope regions of a wide variety of pathogens, including viruses, bacteria, parasites, and microbes. Non-infectious antigens and allergens of interest can also be screened as described herein. Immunization, therapeutic and diagnostic applications are also described for the compositions and methods according to the invention.In another embodiment, the invention provides novel methods of identifying a cryptic neutralizing epitope and related vaccines, constructs, and libraries. In some embodiments, these methods use high-throughput formats that are facilitated by in silica or in vitro steps.

    摘要翻译: 本发明涉及筛选免疫原性病毒样颗粒和相关的免疫原性组合物和诊断技术的方法。 在一个实施方案中,本发明提供筛选含有对应于多种病原体(包括病毒,细菌,寄生虫和微生物)的表位区域的肽的免疫原性病毒样颗粒的方法。 还可以如本文所述筛选感兴趣的非传染性抗原和过敏原。 还描述了根据本发明的组合物和方法的免疫,治疗和诊断应用。 在另一个实施方案中,本发明提供鉴定隐性中和表位和相关疫苗,构建体和文库的新方法。 在一些实施方案中,这些方法使用通过二氧化硅或体外步骤促进的高通量形式。

    PLASMIDS AND METHODS FOR PEPTIDE DISPLAY AND AFFINITY-SELECTION ON VIRUS-LIKE PARTICLES OF RNA BACTERIOPHAGES
    5.
    发明申请
    PLASMIDS AND METHODS FOR PEPTIDE DISPLAY AND AFFINITY-SELECTION ON VIRUS-LIKE PARTICLES OF RNA BACTERIOPHAGES 有权
    肽类显示的方法和方法及RNA病毒类病毒颗粒的选择

    公开(公告)号:US20120295813A1

    公开(公告)日:2012-11-22

    申请号:US13520057

    申请日:2010-12-31

    摘要: The present invention relates to a system and method for controlling peptide display valency on virus-like particles (VLPs), especially including MS2 VLPs. In this method, large amounts of wild-type and low quantities of single-chain dimer coat proteins may be produced from a single RNA. Valency is controlled in immunogen (vaccine) production by providing a system that allows the production of large amounts of wild-type and low quantities of single-chain dimer coating proteins from a single RNA, allowing facile adjustment of display valency levels on VLPs, especially MS2 VLPS over a wide range, from few than one-on average—to as many as ninety per particle. This facilitates the production of immunogens and vaccines, including VLPs exhibiting low valency. Nucleic acid constructs useful in the expression of virus-like particles are disclosed, comprised of a coat polypeptide of MS2 modified by insertion of a heterologous peptide, wherein the heterologous peptide is displayed on the virus-like particle and encapsidates MS2 niRNA. Nucleic acid constructs are also disclosed which are useful in the expression of virus-like particles comprised of a coat polypeptide of PP7 modified by insertion of a heterologous peptide, wherein the heterologous peptide is displayed on the virus-like particle and encapsidates PP7 mRNA.

    摘要翻译: 本发明涉及一种用于控制病毒样颗粒(VLPs),特别是包括MS2 VLP的肽显示效价的系统和方法。 在该方法中,可以从单个RNA产生大量野生型和低量的单链二聚体外壳蛋白。 通过提供允许从单个RNA产生大量野生型和少量单链二聚体包被蛋白质的系统,允许轻度调整VLP上的显示剂价位水平,特别是 MS2 VLPS在广泛的范围内,从一个平均至少一个到每个粒子多达九十个。 这有助于免疫原和疫苗的生产,包括显示低效价的VLP。 公开了可用于病毒样颗粒表达的核酸构建物,其由通过插入异源肽修饰的MS2的外壳多肽组成,其中异源肽显示在病毒样颗粒上并包裹MS2nRNA。 还公开了核酸构建体,其可用于由通过插入异源肽修饰的PP7的外壳多肽组成的病毒样颗粒的表达,其中异源肽显示在病毒样颗粒上并包裹PP7 mRNA。

    Immunogenic HPV L2-Containing VLPs and Related Compositions, Constructs, and Therapeutic Methods
    8.
    发明申请
    Immunogenic HPV L2-Containing VLPs and Related Compositions, Constructs, and Therapeutic Methods 有权
    含免疫原性HPV L2的VLP及相关组合物,构建体和治疗方法

    公开(公告)号:US20140105924A1

    公开(公告)日:2014-04-17

    申请号:US13946562

    申请日:2013-07-19

    IPC分类号: A61K39/12

    摘要: The invention provides immunotherapeutic and prophylactic bacteriophage viral-like particle (VLPs) which are useful in the treatment and prevention of human papillomavirus (HPV) infections and related disorders, including cervical cancer and persistent infections associated with HPV. Related compositions (e.g. vaccines), nucleic acid constructs, and therapeutic methods are also provided. VLPs and related compositions of the invention induce high titer antibody responses against HPV L2 and protect against HPV challenge in vivo. VLPs, VLP-containing compositions, and therapeutic methods of the invention induce an immunogenic response against HPV infection, confer immunity against HPV infection, protect against HPV infection, and reduce the likelihood of infection by HPV infection.

    摘要翻译: 本发明提供免疫治疗和预防性噬菌体病毒样颗粒(VLP),其可用于治疗和预防人乳头状瘤病毒(HPV)感染和相关疾病,包括宫颈癌和与HPV相关的持续感染。 还提供了相关组合物(例如疫苗),核酸构建体和治疗方法。 本发明的VLP和相关组合物诱导针对HPV L2的高滴度抗体应答,并在体内保护免受HPV挑战。 VLP,含VLP的组合物和本发明的治疗方法诱导针对HPV感染的免疫原性应答,赋予HPV感染免疫,防止HPV感染,并降低HPV感染感染的可能性。