ScFv Piezoimmunosensor Detection of Therapeutic Antibodies in Human Serum

ScFv 压电免疫传感器检测人血清中的治疗性抗体

基本信息

批准号：
7788377
负责人：
XIANGQUN ZENG
金额：
$ 23.16万
依托单位：
OAKLAND UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7788377
关键词：
Agonist Antibodies Antigens Autoimmune Diseases Avastin Biological Biological Assay Biological Markers Biotin CYP1B1 gene Cardiovascular Diseases Cetuximab Clinic Clinical Clinical Research Collaborations Coupling Decision Making Detection Development Diagnostic Disease Drops Engineering Ensure Enzymes Erbitux Exhibits Future Generic Drugs Graft Rejection Half-Life Health Care Costs Human Hypersensitivity Immune response Immunoassay Immunoglobulin Fragments Immunologist Industry Injection of therapeutic agent Investigation Joints Label Legal patent Libraries Life Link Malignant Neoplasms Manuscripts Monitor One-Step dentin bonding system Patients Phage Display Physicians Process Protocols documentation Publications Quartz Reagent Recombinant Antibody Recombinants Research Rheumatoid Arthritis Roche brand of rituximab Roche brand of trastuzumab Sampling Serum Specificity Surface Technology Therapeutic Therapeutic Effect Therapeutic Monoclonal Antibodies Therapeutic antibodies Transducers Trastuzumab Work assay development base bevacizumab cell growth cost dosage human disease immunogenic instrumentation killings member neoplastic cell panitumumab point of care public health relevance rituximab success

项目摘要

DESCRIPTION (provided by applicant): Therapeutic monoclonal antibodies (McAbs) exhibit tremendous potential to treat human disease such as cancer and rheumatoid arthritis. More than 300 antibodies are undergoing clinical development and 2915 clinical studies involving antibodies are being carried out. The effect of therapeutic antibodies is concentration dependent. However, in some patients, therapeutic antibodies can be rapidly eliminated from the body and will be of little benefit to the patient. If a patient could have benefited from, but was not given an alternative therapeutic, then the patient's disease could have worsened or become life threatening during the course of therapeutic antibody treatment. However, if a physician can use the results of an assay to readily determine therapeutic antibody concentration, then the physician can adjust treatment to the benefit of the patient. Traditional immunoassays (e.g. enzyme-linked immunosorbant assays or ELISAs) typically incorporate antigens to detect antigen-specific antibodies in serum samples. However, the antigens recognized by many therapeutic antibodies can be unstable, difficult to work with or expensive to acquire. As such, they cannot always be used to build a simple, stable, reproducible assay. Therapeutic antibodies have been engineered or developed to assume most, if not all of the immunological features of a normal human antibody, so that they can be used in humans without eliciting an adverse immune response. Due to their similarities to normal human antibodies, they can be difficult to detect and quantify in human serum without the use of complementary antigens. At present, rapid, simple, highly sensitive, inexpensive assays to detect most of these therapeutic antibodies in human serum samples are not readily available. We have successfully developed a very simple, but extremely powerful scFv-based piezoimmunosensor assay platform to detect unique antibodies in serum samples. We propose in Aim 1 of this project to utilize a large (~2.9 billion member) phage-displayed antibody library to select for stable scFv recombinant antibodies specific for unique therapeutic antibodies. The therapeutic antibody-specific scFv will be used in lieu of traditional antigens for Aim 2 to develop piezoimmunosensors to detect and quantify therapeutic antibodies in human serum samples. The piezoimmunosensors will combine the high specificity and stability of scFv recognition of antigen (i.e. therapeutic antibody) with the low cost, one-step readout of non-label Quartz Crystal Microbalance transducers. Dr. Zeng, an analytical chemist, and Dr. Mernaugh, an immunologist, will carry out the proposed studies. Through their synergistic efforts, this team will create and certify a fast, accurate, highly sensitive and specific, stable, low cost diagnostic assay platform that can be used to detect and quantify therapeutic antibodies (e.g. Bevacizumab, Cetuximab, Panitumumab and Trastuzumab) in human serum samples. PUBLIC HEALTH RELEVANCE: The proposed research will focus on developing scFv recombinant antibodies and scFv-based piezoimmunosensor assays for use in detecting therapeutic antibodies in human serum samples. At present, rapid, simple, highly sensitive, inexpensive assays to detect most of therapeutic antibodies in human serum samples are not readily available. Since the proposed assay will be more sensitive than traditional immunoassay (e.g. ELISAs or whole antibody immunosensors) and can rapidly and accurately detect therapeutic antibodies in humans using a few drops of human serum and inexpensive instrumentation, a physician will be able to quickly determine if the concentration of therapeutic antibody in patients is sufficient or if therapy must be changed to benefit the patient.

描述（由申请人提供）：治疗性单克隆抗体（McAb）在治疗癌症和类风湿性关节炎等人类疾病方面表现出巨大的潜力。超过300种抗体正在进行临床开发，2915项涉及抗体的临床研究正在进行中。治疗性抗体的作用是浓度依赖性的。然而，在某些患者中，治疗性抗体可以迅速从体内消除，对患者没有什么好处。如果患者本可以受益，但没有给予替代疗法，那么在治疗性抗体治疗过程中，患者的疾病可能会恶化或危及生命。然而，如果医生可以使用测定结果来容易地确定治疗性抗体浓度，则医生可以调整治疗以使患者受益。传统的免疫测定（例如酶联免疫吸附测定或 ELISA）通常结合抗原来检测血清样本中的抗原特异性抗体。然而，许多治疗性抗体识别的抗原可能不稳定、难以使用或获取成本昂贵。因此，它们不能总是用于构建简单、稳定、可重复的测定。治疗性抗体经过工程设计或开发，具有正常人类抗体的大部分（如果不是全部）免疫学特征，因此它们可以用于人类而不会引起不良免疫反应。由于它们与正常人类抗体相似，因此在不使用互补抗原的情况下很难在人血清中检测和定量它们。目前，用于检测人血清样品中大多数这些治疗性抗体的快速、简单、高灵敏度、廉价的测定方法尚不可用。我们成功开发了一种非常简单但功能极其强大的基于 scFv 的压电免疫传感器检测平台，用于检测血清样本中的独特抗体。我们在该项目的目标 1 中建议利用大型（约 29 亿成员）噬菌体展示抗体库来选择对独特治疗抗体具有特异性的稳定 scFv 重组抗体。 Aim 2 将使用治疗性抗体特异性 scFv 代替传统抗原，开发压电免疫传感器来检测和量化人血清样本中的治疗性抗体。压电免疫传感器将 scFv 识别抗原（即治疗性抗体）的高特异性和稳定性与非标记石英晶体微天平传感器的低成本、一步读出相结合。分析化学家 Zeng 博士和免疫学家 Mernaugh 博士将开展拟议的研究。通过他们的协同努力，该团队将创建并认证一个快速、准确、高灵敏度、特异、稳定、低成本的诊断测定平台，可用于检测和量化人类治疗抗体（例如贝伐单抗、西妥昔单抗、帕尼单抗和曲妥珠单抗）血清样本。公共健康相关性：拟议的研究将重点开发 scFv 重组抗体和基于 scFv 的压电免疫传感器测定法，用于检测人血清样本中的治疗抗体。目前，用于检测人血清样品中大多数治疗性抗体的快速、简单、高灵敏度、廉价的检测方法尚不可用。由于所提出的测定法比传统的免疫测定法（例如 ELISA 或全抗体免疫传感器）更灵敏，并且可以使用几滴人血清和廉价仪器快速准确地检测人体中的治疗性抗体，因此医生将能够快速确定是否存在患者体内的治疗抗体浓度是否足够，或者是否必须改变治疗以使患者受益。