Liquid Chromatography System for the purification of peptidic radiotracers, bioconjugates, nucleosides, and drug-like molecules

用于纯化肽放射性示踪剂、生物缀合物、核苷和类药物分子的液相色谱系统

• 批准号: RTI-2019-00708
• 负责人: Perrin, David
• 金额: $ 7.6万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=661986
• 关键词: Liquid; Chromatography; System; purification; peptidic

Training: My trainees learn to synthesize novel monomer amino acids, nucleosides, boronic acids and incorporate these groups into complex bioconjugates (peptides and oligonucleotides) that will be used for cancer detection, diagnosis and treatment. Trainees in collaborators lab's will purify peptides for metal-mediated catalysis and resolve racemic mixtures of novel amino acids.******Indispensable Equipment: an HPLC (high performance liquid chromatograph) with diode-array UV-vis and auto-sampling is essential to purify and demonstrate purity for publication. We must replace/upgrade an aging 13-year-old HPLC to enable my trainees in my lab to produce key molecules for research, in collaboration with PIs at B.C. Cancer Research Agency, UCLA, and at local companies, Zymeworks Inc. and Iprogen. Trainees acquire skills in synthetic organic, bioconjugate, radio- and biological chemistry as they develop novel probes to translate these novel constructs into applications for human health. Over the past 13 years, we have established a strong and diverse publication record along with several patents. Specifically, trainees work on: 1) a proprietary class of peptide-trifluoroborate conjugates for PET imaging; 2) synthetically modified DNAs for catalysis and target binding that depend on modified nucleoside triphosphates, and 3) the total synthesis of amanitin, a toxic peptide natural product and “clickable” analogs for antibody-drug conjugates. Hence an HPLC is as essential for my lab. ******Required for Publication: In order to publish these results, an HPLC with detection hardware is absolutely required; chromatograms backed up by advanced software analysis based on diode-array multi-wavelength detection give confidence to collaborators for translation. Because we work with radioactive molecules (32P), safety regulations necessitate a dedicated HPLC within a lab like mine, that is licensed for radioisotope use. ******Dependent Collaborations: In addition, increased funding from the CIHR and the CCSRI (which do not allow for major equipment purchases) fund an increased number of trainees, which in turn have placed severe constraints on the machine, which runs nearly 24/7 via autosampler. Collaborations with Paul Schaffer (Triumf), and Kuo-Shyan Lin (BC Cancer Research Centre), Laurel Schafer and Jason Hein (UBC Chemistry) place my lab as a hub for generating numerous molecules that interest other key players at UBC. Finally, my lab's productivity is now severely jeopardized by a 13-year old HPLC that routinely fails, requiring part replacements, in many cases on the used market since the model is so old that Agilent no longer makes all the parts. Outdated software will be threatened by the need to replace the computer that services the HPLC yet where the software is not compatible with the new Windows operating system. ******Summary: a new HPLC is requested to support a highly productive and collaborative research program at the interface of chemistry and biology. **

培训内容：我的学员学习合成新的单体氨基酸，核苷，硼酸，并将这些基团纳入复杂的生物缀合物（肽和寡核苷酸），将用于癌症检测，诊断和治疗。合作者实验室的实习生将纯化用于金属介导催化的肽，并解析新型氨基酸的外消旋混合物。仪器设备：具有二极管阵列UV-vis和自动取样的HPLC（高效液相色谱仪）对于纯化和证明出版物的纯度是必要的。我们必须更换/升级老化的13岁的HPLC，使我的实习生在我的实验室生产的关键分子的研究，与PI在不列颠哥伦比亚省癌症研究机构，加州大学洛杉矶分校，并在当地公司，Zymeworks公司。还有伊普罗金受训者获得合成有机，生物共轭，无线电和生物化学的技能，因为他们开发新的探针，将这些新的结构转化为人类健康的应用。在过去的13年里，我们已经建立了一个强大的和多样化的出版记录沿着与几项专利。具体来说，受训人员从事：1）用于PET成像的专有肽-三氟硼酸盐缀合物; 2）用于催化和靶向结合的合成修饰的DNA，其依赖于修饰的核苷三磷酸;和3）鹅膏蕈碱的全合成，一种有毒肽天然产物和抗体药物缀合物的“可点击”类似物。因此，HPLC对我的实验室至关重要。** 发表要求：为了发表这些结果，绝对需要配备检测硬件的HPLC;基于二极管阵列多波长检测的高级软件分析支持的色谱图为合作者提供了翻译的信心。因为我们使用放射性分子（32 P），安全法规要求在像我这样的实验室中使用专用的HPLC，该实验室被许可使用放射性同位素。** 相关合作：此外，CIHR和CCSRI（不允许购买主要设备）增加的资金为更多的学员提供资金，这反过来又对机器造成了严重的限制，该机器通过自动采样器运行近24/7。与Paul Schaffer（Triumf）和Kuo-Shyan Lin（BC癌症研究中心），月桂Schafer和Jason Hein（UBC化学）的合作使我的实验室成为产生UBC其他关键参与者感兴趣的众多分子的中心。最后，我的实验室的生产力现在严重危及一个13岁的HPLC，经常失败，需要更换部件，在许多情况下，在二手市场上，因为该模型是如此之旧，安捷伦不再使所有的部分。过时的软件将受到威胁，因为需要更换为HPLC提供服务的计算机，但该软件与新的Windows操作系统不兼容。** 总结：需要一种新的HPLC来支持化学和生物学界面上的高效协作研究项目。**