Development of Room-Temperature Storage Technique for Plasma/Serum Biospecimens

血浆/血清生物样本室温储存技术的发展

• 批准号: 8546307
• 负责人: Alptekin Aksan
• 金额: $ 17.61万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-17 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8546307
• 关键词: Adsorption; Ascites; Biochemical Reaction; Biological Markers; Biological Preservation; Bronchoalveolar Lavage Fluid; Characteristics; Chemicals; Cold Chains; Collection; DNA; Desiccation; Development; Diagnostic; Epidemiology; Equipment; Farming environment; Freezing; Future; Genomics; Glass; Goals; Hour; Hydration status; Ice; Kinetics; Lipids; Liquid substance; Malignant Neoplasms; Medicine; Methodology; Methods; Outcome; Paper; Plasma; Porifera; Process; Proteins; Proteomics; Protocols documentation; RNA; Research; Research Project Grants; Saliva; Sampling; Serum; Serum Proteins; Site; Specimen; Stress; Structure; Techniques; Technology; Temperature; Testing; Therapeutic; Time; United States National Institutes of Health; Urine; Weight; biobank; cost; cryogenics; experience; improved; innovation; macromolecule; metabolomics; novel; research study; success; sugar; tool

DESCRIPTION (provided by applicant): Currently, millions of serum biospecimens are being stored in biorepositories across the nation, while tens of thousands of new biospecimens are added to the pool daily. These biospecimens are stored for future research, mainly for proteinaceous biomarker discovery and verification (e.g. for diagnostic, therapeutic, and epidemiologic outcomes). The success of biomarker research not only depends upon the availability of the tools (proteomic, peptidomic, lipidomic and metabolomic technologies) to extract information from biospecimens, but also on the availability of "high quality" biospecimens. However, in most biorepositories, serum is stored by freezing without following any preservation protocol; the samples are directly placed in -20, -40 or -80oC freezers, in the absence of any cryoprotectant, where they experience very slow cooling (1-2o C/min). It is well known that these conditions impose very harsh chemical and physical stresses on macromolecules, altering their characteristics (structure and activity), often irreversibly. Recent evidence has shown that some of the most promising proteinaceous cancer biomarkers are indeed very susceptible to freeze/thaw and frozen state storage. Therefore, it is plausible that frozen state storage may cause most of the potential biomarker information in the stored sera biospecimens to be lost forever. Our long-term goal is to eliminate the requirement for frozen state storage and develop the techniques to store serum biospecimens at room temperature using isothermal vitrification technology. Isothermal vitrification is the process by which liquids doped with sugars are desiccated to a "glass" (a very viscous fluid). In this state, biochemical reactions are halted, degradation of the specimen is stopped, and macromolecules are stabilized in their native states. Isothermal vitrification will eliminate the exposure of the proteinaceous biomarkers to freeze/thaw stresses and to frozen state storage damage and thus will substantially increase the quality of the stored biospecimens. It will also present a more economical alternative to freezing, since storage of specimens in freezer-farms will no longer be needed. We will accomplish our goal by achieving the following four Aims: Aim #1. Develop a panel of lyoprotectant chemicals to be added to serum samples for isothermal vitrification: Demonstrate that the lyoprotectant cocktail enables rapid and uniform vitrification while stabilizing sera proteins. Aim #2. Determine the retention and elution efficiency of sera proteins following vitrification by adsorption in standard filter paper using the developed lyoprotectant cocktail. Aim #3. Determine whether specific sera proteins are altered following optimized vitrification and elution conditions from the optimal matrix; document protein depletion, aggregation and degradation. Aim #4. Develop and validate an electrospun sponge for streamlined vitrification of serum samples at sera collection sites.

描述（由申请人提供）：目前，全国各地的生物库中储存着数百万份血清生物标本，每天有数以万计的新生物标本加入。这些生物标本被保存以备将来研究，主要用于蛋白质生物标志物的发现和验证（例如用于诊断、治疗和流行病学结果）。生物标志物研究的成功不仅取决于从生物标本中提取信息的工具（蛋白质组学、肽组学、脂质组学和代谢组学技术）的可用性，还取决于“高质量”生物标本的可用性。然而，在大多数生物储存库中，血清是通过冷冻储存的，没有遵循任何保存方案；在没有任何冷冻保护剂的情况下，样品直接放置在-20、-40或-80℃的冰柜中，在那里它们经历非常缓慢的冷却（1 -20℃/分钟）。众所周知，这些条件对大分子施加了非常严酷的化学和物理压力，常常不可逆转地改变了它们的特性（结构和活性）。最近