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

• 批准号: 8010212
• 负责人: XIANGQUN ZENG
• 金额: $ 18.84万
• 依托单位: OAKLAND UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8010212
• 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; Legal patent; Libraries; Life; Link; Malignant Neoplasms; Manuscripts; Monitor; Monoclonal Antibodies; 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

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）在治疗人类疾病如癌症和类风湿性关节炎方面表现出巨大的潜力。300多种抗体正在进行临床开发，2915项涉及抗体的临床研究正在开展。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，对患者几乎没有好处。如果患者可以受益于替代治疗，但没有给予替代治疗，则患者的疾病可能在治疗性抗体治疗过程中恶化或变得危及生命。然而，如果医生可以使用测定的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）通常掺入抗原以检测血清样品中的抗原特异性抗体。然而，被许多治疗性抗体识别的抗原可能是不稳定的，难以使用或获取昂贵。因此，它们不能总是用于建立简单、稳定、可重复的测定。治疗性抗体已经被工程化或开发以呈现正常人抗体的大多数（如果不是全部的话）免疫学特征，使得它们可以用于人而不引发不利的免疫应答。由于它们与正常人抗体的相似性，在不使用互补抗原的情况下，它们可能难以在人血清中检测和定量。目前，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、廉价的测定法还不容易获得。 我们已经成功地开发了一个非常简单，但非常强大的基于scFv的压电免疫传感器检测平台，以检测血清样品中的独特抗体。我们在该项目的目的1中提出利用大的（~ 29亿成员）噬菌体展示抗体文库来选择对独特的治疗性抗体特异性的稳定的scFv重组抗体。治疗性抗体特异性scFv将用于替代Aim 2的传统抗原，以开发压电免疫传感器来检测和定量人血清样品中的治疗性抗体。压电免疫传感器将联合收割机抗原（即治疗性抗体）的scFv识别的高特异性和稳定性与非标记石英晶体微天平换能器的低成本、一步读出。 分析化学家Zeng博士和免疫学家Mernaugh博士将进行拟议的研究。通过他们的协同努力，该团队将创建并认证一个快速，准确，高灵敏度和特异性，稳定，低成本的诊断分析平台，可用于检测和定量人血清样品中的治疗性抗体（例如贝伐单抗，西妥昔单抗，帕尼单抗和曲妥珠单抗）。 公共卫生相关性：拟议的研究将集中于开发scFv重组抗体和基于scFv的压电免疫传感器测定用于检测人血清样品中的治疗性抗体。目前，快速、简便、高灵敏度、低成本的检测人血清中大多数治疗性抗体的方法尚不容易获得。由于所提出的测定将比传统的免疫测定（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的仪器快速且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患者中的治疗性抗体的浓度是否足够或者是否必须改变治疗以使患者受益。

项目成果

Glycosylated aniline polymer sensor: amine to imine conversion on protein-carbohydrate binding.

• DOI: 10.1016/j.bios.2013.02.030
• 发表时间: 2013-08-15
• 期刊: BIOSENSORS & BIOELECTRONICS
• 影响因子: 12.6
• 作者: Wang, Zhe;Sun, Chunyan;Vegesna, Giri;Liu, Haiying;Liu, Yang;Li, Jinghong;Zeng, Xiangqun
• 通讯作者: Zeng, Xiangqun

Real time analysis of binding between Rituximab (anti-CD20 antibody) and B lymphoma cells.

• DOI: 10.1021/ac400062v
• 发表时间: 2013-09-17
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Tan, Liang;Lin, Peiling;Chisti, Mohammad M.;Rehman, Abdul;Zeng, Xiangqun
• 通讯作者: Zeng, Xiangqun

Immobilization of a human epidermal growth factor receptor 2 mimotope-derived synthetic peptide on Au and its potential application for detection of herceptin in human serum by quartz crystal microbalance.

• DOI: 10.1021/ac201430p
• 发表时间: 2011-12-01
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Shang, Yuqin;Singh, Pankaj R.;Chisti, Mohammad M.;Mernaugh, Ray;Zeng, Xiangqun
• 通讯作者: Zeng, Xiangqun

Characterization of the native and denatured herceptin by enzyme linked immunosorbent assay and quartz crystal microbalance using a high-affinity single chain fragment variable recombinant antibody.

通过酶连接的免疫吸附测定法和石英晶体微生物的酶来表征天然和变性的赫赛汀，使用高亲和力的单链片段可变重组抗体。

• DOI: 10.1021/ac301235a
• 发表时间: 2012-10-02
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Shang, Yuqin;Mernaugh, Ray;Zeng, Xiangqun
• 通讯作者: Zeng, Xiangqun

Real-time monitoring of cell mechanical changes induced by endothelial cell activation and their subsequent binding with leukemic cell lines.

• DOI: 10.1016/j.bios.2014.01.004
• 发表时间: 2014-06-15
• 期刊: BIOSENSORS & BIOELECTRONICS
• 影响因子: 12.6
• 作者: Tan, Liang;Lin, Peiling;Pezeshkian, Bahareh;Rehman, Abdul;Madlambayan, Gerard;Zeng, Xiangqun
• 通讯作者: Zeng, Xiangqun