DEVELOPMENT OF A QUANTITATIVE ANTIBODY BINDING KIT

各种疾病的诊断（例如，白血病和艾滋病） 可以通过研究不同的亚群来促进 血液中的白细胞 尽管单克隆抗体具有 已成功用于定性表征许多 病理性疾病，它仍然非常难以量化 在特定细胞类型上的单克隆抗体结合位点。 抗体结合位点数目的变化可作为抗体结合位点数目的变化。 疾病阶段的指数，以及 特殊医疗的有效性。 的目的 建议是为临床流程开发校准套件 允许一种简单、直接和廉价方法的细胞计数器 为了确定每个单克隆抗体结合位点的数量， cell. 该方法将基于一组高度统一的 微珠，其在大小和大小方面表现为“模型细胞”， 抗体结合行为 这些微珠会结合特定的 荧光单克隆小鼠 IgG抗体，并将作为标准构建 仪器响应与 使用流式细胞仪检测荧光抗体。 在第一阶段， 项目，它将被证明，这个校准图可能是 构建并用于确定抗体结合的数量 特定细胞群的位置。 项目第二阶段将 与临床医生密切合作，评估临床 该试剂盒的适用性及其规模化生产 和化学化。 预计这项新技术 将为临床和研究实验室提供独特的工具， 对某些疾病的诊断、研究和治疗。

The diagnosis of a variety of diseases (e.g., leukemia and AIDS) can be facilitated by studying the various sub-populations of leukocytes in the blood. Although monoclonal antibodies have been used successfully to qualitatively characterize many pathological diseases, it remains extremely difficult to quantitate monoclonal antibody binding sites on a particular cell type. Changes in the number of antibody-binding sites may serve as an index to the stage of a disease, as well as, an indicator of the effectiveness of particular medical treatments. The aim of this proposal is to develop a calibration Kit for the clinical flow cytometer that will allow an easy, direct, and inexpensive method to determine the number of monoclonal antibody binding sites per cell. The method will be based on a set of highly uniform microbeads, that behave as "model cells" in terms of both size and antibody binding behavior. These microbeads will bind specific amounts of commerically available fluorescent monoclonal mouse IgG antibodies and will serve as the standards to construct calibration plots of instrument response against the number of fluorescent antibodies using a flow cytometer. In Phase I of this project, it will be demonstrated that this calibration plot may be constructed and used to determine the number of antibody binding sites for particular cell populations. Phase II of the project will involve close collaboration with clinicians to assess the clinical applicability of this Kit, as well as, its scale-up manufacturing and commericalization. It is expected that this new technology will provide clinical and research laboratories with unique tool in the diagnosis, study, and treatment of certain disorders.