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药物，但其高昂的成本使其成为假冒或非法分销的诱人目标。在美国和世界范围内都发生了许多病例，药品分销网络的复杂性使得这个问题可能会持续存在。该项目的目标是开发一种简单且廉价的横向流动免疫分析法，可用于快速确定单克隆抗体治疗的完整性。我们的测试使用模拟单抗识别的天然抗原表位的拟态肽，可以区分活性单抗和其他蛋白质或免疫球蛋白污染物。Mimetope肽已被开发用于肿瘤学领域的前三大单抗：贝伐单抗、曲妥珠单抗和利妥昔单抗。利妥昔单抗mimotopes已经以ELISA格式开发并用于药代动力学研究，其余两个将在本提案的Aim 1中进行类似的验证。在目标2中，我们将把ELISA分析转化为侧流免疫分析格式。在Aim 3中，我们将与Moores UCSD癌症中心的一位顶级临床科学家合作，对每个单抗的原型测试进行双盲研究，确定它们的准确性以及它们检测生理盐水或多克隆人类免疫球蛋白的能力。这些产品的成功开发将使肿瘤学家和他们的患者对这些潜在的挽救生命的治疗方法在给药时是真实和有效的增加信心。

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.