DEVELOPMENT OF A QUANTITATIVE ANTIBODY BINDING KIT

多种疾病的诊断(如白血病和艾滋病) 可以通过研究不同的亚种群来促进 血液中的白细胞。尽管单抗具有 被成功地用来定性地描述了许多 病理性疾病，仍然极难量化 一种特定细胞类型上的单抗结合部位。 抗体结合位点数的变化可以作为一种 疾病分期的指标，也是疾病发展的指标 特定医疗方法的有效性。这样做的目的是 建议开发用于临床流程的校准试剂盒 这将是一种简单、直接和廉价的方法 以确定每个单抗结合位点数。 手机。该方法将基于一套高度统一的 微珠，在大小和大小方面都表现为“模型细胞” 抗体结合行为。这些微珠将结合特定的 商品化荧光单克隆鼠的数量 并将作为构建的标准 仪器响应相对于数量的校准曲线图 用流式细胞仪检测荧光抗体。在这个项目的第一阶段 项目，它将证明这个校准图可能是 构建并用于确定抗体结合数 特定细胞群体的地点。该项目的第二阶段将 与临床医生密切合作，以评估临床 该套件的适用性以及它的扩大制造 和商业化。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.