Rapid and Inexpensive Characterisation of Immunoassay Conjugates (RIChIC)

快速且廉价地表征免疫分析偶联物 (RIChIC)

• 批准号: 105569
• 金额: $ 4.46万
• 依托单位: FLEET BIOPROCESSING LIMITED
• 依托单位国家: 英国
• 项目类别: Collaborative R&D
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=105569
• 关键词: Rapid; Inexpensive; Characterisation; Immunoassay; Conjugates

Fleet Bioprocessing Ltd. are experts in the development of immunoassays, widely-used tests for the diagnosis of diseases which rely on the well-known specificity of antibodies to detect specific molecules. Examples in common use include tests for detecting HIV and hepatitis, for diagnosing thyroid hormone abnormalities, or for differentiating heart attacks from other conditions such as angina which may display similar symptoms. New immunoassays are under development all the time, e.g. to improve the detection of cancer tumours or to monitor factors associated with the development of Alzheimer's disease.Immunoassays rely on the successful chemical "labelling" of antibodies and related proteins, so for example that they can be detected efficiently via the presence of a fluorescent dye - and Fleet are expert in the bioconjugation techniques required for this purpose. Fleet routinely use simple analytical techniques to characterise these labelled antibody conjugates, allowing us to determine basic information such as the antibody concentration and the mean number of dye molecules per antibody molecule.However these techniques tell us nothing about whether the labelled antibody conjugate has retained its ability to detect the molecule of interest, or has been damaged in the labelling process. For example it is possible to attach too many dye molecules to an antibody, with the result that its ability to bind the target molecule is compromised. It would be very useful to have access to a rapid, inexpensive analytical method allowing us to confirm that the conjugate has successfully retained its structure during the labelling procedure.Fleet have evaluated several techniques for this purpose, but to date all have failed to meet our requirements; simple techniques based on spectroscopy which would meet our needs of being rapid and inexpensive have not shown adequate sensitivity to differentiate between "good" and "bad" conjugates, while techniques capable of achieving the required sensitivity have proved prohibitively expensive and/or time-consuming.An initial project with LGC and NPL (completed in March 2019) identified techniques with exciting potential to fill this knowledge gap, and to better understand the mechanism of conjugate inactivation. This follow-up project aims to confirm the potential of these techniques. Fleet will prepare a range of antibody conjugates for evaluation and assess them in a model immunoassay, while LGC and NPL will characterise them using a range of candidate analytical techniques. This will hopefully allow us to confirm the capability of these techniques for routine use by Fleet Bioprocessing Ltd.

Fleet Bioprocessing Ltd.是开发免疫分析的专家，这种广泛用于疾病诊断的测试依赖于众所周知的抗体特异性来检测特定分子。常用的例子包括检测艾滋病毒和肝炎，诊断甲状腺激素异常，或将心脏病发作与可能表现出类似症状的心绞痛等其他病症区分开来。新的免疫测定法一直在开发中，例如，改进癌症肿瘤的检测或监测与阿尔茨海默病发展有关的因素。免疫测定依赖于抗体和相关蛋白质的成功化学“标记”，例如，可以通过荧光染料的存在有效地检测它们- Fleet是为此目的所需的生物偶联技术方面的专家。Fleet常规使用简单的分析技术来表征这些标记的抗体偶联物，使我们能够确定基本信息，如抗体浓度和每个抗体分子的平均染料分子数。然而，这些技术并没有告诉我们标记的抗体偶联物是否保留了检测感兴趣分子的能力，或者在标记过程中已经损坏。例如，有可能在抗体上附着太多的染料分子，其结果是其结合目标分子的能力受到损害。这将是非常有用的，有机会获得快速，廉价的分析方法，使我们能够确认在标记过程中成功地保留其结构的共轭物。Fleet为此目的评估了几种技术，但迄今为止，所有技术都未能满足我们的要求；基于光谱学的简单技术可以满足我们快速和廉价的需求，但却没有显示出足够的灵敏度来区分“好”和“坏”共轭物，而能够达到所需灵敏度的技术已被证明过于昂贵和/或耗时。LGC和NPL的初始项目（于2019年3月完成）确定了具有令人兴奋的潜力的技术，以填补这一知识空白，并更好地了解共轭失活的机制。这个后续项目旨在确认这些技术的潜力。Fleet将准备一系列抗体偶联物用于评估，并在模型免疫分析中对其进行评估，而LGC和NPL将使用一系列候选分析技术对其进行表征。这将使我们有希望确认这些技术在Fleet Bioprocessing有限公司的日常使用能力。