Lateral Flow Immunoassay for Therapeutic Monoclonal Antibody Quality Assurance

描述（由申请人提供）：单克隆抗体（mAb）是治疗癌症和其他疾病的有价值的治疗剂，但其高成本使其成为假冒或非法分销的诱人目标。美国和世界各地都发生了许多病例，药品分销网络的复杂性使问题可能持续存在。该项目的目标是开发一种简单而廉价的侧流免疫测定法，可用于快速确定单克隆抗体治疗剂的完整性。使用模拟给定mAb识别的天然抗原表位的模拟肽，我们的测试将区分活性mAb和其他蛋白质或免疫球蛋白污染物。已经开发了用于肿瘤学中使用的前三种mAb的MIPs肽：贝伐珠单抗、曲妥珠单抗和利妥昔单抗。利妥昔单抗模拟表位已经以ELISA形式开发并用于药代动力学研究，其余两种将在本提案的目标1中进行类似验证。在目标2中，我们将ELISA测定转化为侧流免疫测定形式。在目标3中，我们将与Moores UCSD癌症中心的顶级临床医生-科学家合作，对每种mAb的原型测试进行双盲研究，确定其准确性以及检测生理盐水或多克隆人免疫球蛋白的能力。这些产品的成功开发将使肿瘤学家及其患者更加相信这些潜在的挽救生命的疗法在给药时是真实且有效的。

DESCRIPTION (provided by applicant): Monoclonal antibodies (mAb) are valuable therapeutics for treating cancer and other diseases, but their high cost makes them attractive targets for counterfeiting or illegal distribution. Numerous cases have occurred both in the US and worldwide and the complexity of the pharmaceutical distribution network makes the problem likely to persist. The goal of this project is to develop a simple and inexpensive lateral flow immunoassay that can be used to quickly determine the integrity of a monoclonal antibody therapeutic. Using mimetope peptides that mimic the native antigen epitope recognized by a give mAb, our test will discriminate between active mAb and other protein or immunoglobulin contaminants. Mimetope peptides have been developed for the top three mAb used in oncology: bevacizumab, trastuzumab, and rituximab. The rituximab mimetopes have already been developed in an ELISA format and used for pharmacokinetic studies, the remaining two will be similarly validated in Aim 1 of this proposal. In Aim 2, we will translate the ELISA assay to the lateral flow immunoassay format. In Aim 3 we will collaborate with a top clinician-scientist at the Moores UCSD Cancer Center to conduct a double blind study of the prototype tests for each of the mAb, determining their accuracy as well as their ability to detect either saline or polyclonal human immunoglobulin. Successful development of these products will enable oncologists and their patients to have added confidence that these potentially life-saving therapies are genuine and active at the time of administration.