Wilson M R, Wotherspoon J S
Clinical Immunology Research Centre, University of Sydney, N.S.W., Australia.
J Immunol Methods. 1988 Mar 16;107(2):225-30. doi: 10.1016/0022-1759(88)90222-0.
The quantitative and qualitative capacities of flow cytometric analysis that have made it such a powerful tool in studies of cellular antigens have not previously been exploited when dealing with non-cellular antigens. A new immunofluorescence assay technique was developed, using an indirect staining procedure with monoclonal anti-kappa antibodies, to detect human free kappa light chains covalently bound to microspheres of a size suitable for flow cytometry. The strength of the fluorescent signal produced on the microspheres was related to the amount of antigen bound and the size of the beads. At the time of this work large microspheres (i.e., greater than 3 micron in diameter) suitable for this application were only available as suspensions of polysized beads. The fluorescent signal detected on labelled beads was optimized by selecting for analysis, on the basis of the forward angle laser scatter, only those beads of largest diameter. There are many potential applications for this technique - microspheres can be used for the presentation of virtually any antigen or antibody. The analytical benefits inherent in flow cytometry would be a significant advantage in the development of quantitative assays using this method.
流式细胞术分析的定量和定性能力使其成为细胞抗原研究中的强大工具，但在处理非细胞抗原时，这些能力此前尚未得到利用。我们开发了一种新的免疫荧光检测技术，采用间接染色程序和单克隆抗κ抗体，以检测与适合流式细胞术的大小的微球共价结合的人游离κ轻链。微球上产生的荧光信号强度与结合的抗原量和珠子大小有关。在这项工作开展时，适合此应用的大微球（即直径大于3微米）仅以多尺寸珠子的悬浮液形式提供。通过基于前向角激光散射选择仅分析最大直径的那些珠子，优化了在标记珠子上检测到的荧光信号。该技术有许多潜在应用——微球可用于呈现几乎任何抗原或抗体。流式细胞术固有的分析优势在使用该方法开发定量测定中将是一个显著优势。
