Silk-Based Blood Sample Stabilization for Prostate Cancer Diagnostics

 说明书(由申请人提供)迫切需要一种血样制备和处理方法，该方法可以保持完整性并改进下游分析，同时完全不需要冷藏要求。这种技术将允许临床医生和患者舒适地从患者家中收集血液样本进行癌症筛查、监测和/或诊断，从而消除了去门诊实验室的需要。Vaxess Technologies正在开发一套基于生物材料的方法，使用一种新型的丝蛋白基质来提高各种样品的稳定性。这项建议的目的是表征丝素基质稳定血液样本中一组代谢性前列腺癌生物标志物的能力。基于丝绸的保护的增强性能将与其他类似方法(干血斑(DBS)卡)和黄金标准(冷冻储存)进行比较。我们预计，丝绸的使用将允许最小限度或非侵入性的样品收集途径，并将提供与当前黄金标准冷冻存储相当的分析物保留，同时与商业上可用的DBS卡相比，提高了回收率。Vaxess拥有广泛的初步数据，证明了使用丝素稳定全血、血浆、血清和唾液等复杂液体中的抗体、血浆蛋白质和核酸。我们预计可以对血液分析做同样的事情，就像这里建议的那样。在第一个目标中，我们将形成血清负载的丝素基质并定量测定代谢性前列腺癌标记物的水平。这将通过使用质谱学来测量从丝绸基质中回收分析物，并将产量直接与冷冻对照或DBS卡进行比较来实现。蚕丝分子量和净电荷的修饰可以系统地改变蛋白质组学相互作用，从而为从样品回收的角度研究蚕丝稳定化的机理提供了一条途径。在第二个目标中，我们将比较目标1中确定的标记与冷冻对照样本和DBS卡的稳定性，这些样本都在不同的温度和湿度水平下进行处理。这些研究的预期结果将是开发一种基于丝绸的样品制备方法，用于癌症特异性分析物，这将改善下游样品处理。这项拟议研究的基本原理是，一旦这些生物标志物的恢复和稳定性被证明通过丝素包裹体得到增强，诊断试剂盒就可以配备各种丝素形式，然后可以作为平台系统在后续研究中进行充分验证。计划方法的成功可以通过增加获得精确分析的总体机会和减少影响样本质量的分析前差异，对癌症诊断领域的许多需求领域产生积极影响，包括早期检测、筛查和临床诊断。此外，通过提供与指刺全血体积对接的方法，基于丝绸的稳定系统可以在没有抽血员的情况下使用，从而绕过门诊对资源的要求。

 DESCRIPTION (provided by applicant) There is a critical need for a blood sample preparation and processing method that can preserve integrity and improve downstream analysis, while being completely free of cold storage requirements. Such a technology would allow clinicians and patients to collect blood samples for cancer screening, monitoring and/or diagnosis comfortably from patient's homes, obviating the need for outpatient lab visits. Vaxess Technologies is developing a set of biomaterial-based approaches that improves the stability of a wide range of samples using a novel silk protein matrix. The objective of this proposal is to characterize the ability of a silk matrix to stabilize a panel of metabolomic prostate cancer biomarkers in a blood sample. The enhanced performance of silk- based protection will be compared to other similar methods (dried blood spot (DBS) cards) and the gold standard (frozen storage). We anticipate that the use of silk will allow for minimally- or non-invasive sample collection routes, and will provide comparable analyte retention to the current gold standard, frozen storage, while improving recovery as compared to commercially-available DBS cards. Vaxess has extensive preliminary data demonstrating the use of silk to stabilize antibodies, plasma proteins and nucleic acids from complex fluids such as whole blood, plasma, serum, and saliva. We anticipate being able to do the same with blood analytes, as proposed here. In the first aim, we will form serum-laden silk matrices and quantitate metabolomic prostate cancer marker levels. This will be accomplished by using mass spectrometry to measure analyte recovery from silk matrices and comparing the yields directly to frozen controls or DBS cards. Modifications of silk molecular weight and its net charge can systematically change proteomic interactions, and thus provides a route to study the mechanistic underpinnings of silk stabilization in the context of sample recovery. In a second aim, we will compare the stability of markers identified in Aim 1 to frozen control samples and DBS cards when all have been treated at various temperatures and humidity levels. The expected outcome of these studies will be the development of a silk-based sample preparation method for cancer-specific analytes that will improve downstream sample processing. The rationale for the proposed research is that, once recovery and stability of these biomarkers is proven to be augmented by a silk encapsulate, diagnostic kits can be outfitted with various silk formats, which can then be used as a platform system for full validation in follow-on studies. Success with the planned approach can positively impact many areas of need in the field of cancer diagnostics, including early detection, screening, and clinical diagnosis, by increasing overall access to precise analytics and by reducing pre-analytical variations that affect sample quality. Furthermore, by offering methods to interface with finger-prick volumes of whole blood, a silk-based stabilization system could be used without a phlebotomist and thus circumvent the resource requirements of the outpatient clinic.