异硫氰酸荧光素标记抗体的固体基质室温磷光性质研究

研究了异硫氰酸荧光素（FITC）标记的羊抗人抗体（GAHAb-FITC)和兔抗羊抗体9RAGAb-FITC)，及其以三种免疫方式与人免疫球蛋白（IgG)反应所得免疫复合物在多种固体薄膜基质上发射室温磷光（RTP）的适宜条件及其光谱，强度和寿命等性质。研究发现，标记抗体及其免疫复合物保留了FITC优良的固体基质室温磷光性质，λ m0643em^(max)=525/650nm，其RTP强度与其浓度线性相关，并有很高的灵敏度，更为重要的是，免疫反应及其RTP检测能结合于同一基质，方便地相继完成，在聚酰胺膜（PM）上，以Pb(Ac)2为重原子微扰剂，稀释比为1：10（ψ）的GAHAb-FITC,RAGAb-FITC及其与人IgG形成的免疫复合物均能发射较强制 RTP信号并有较长的RTP寿命，本结果为建立相应的固体基质室温磷光免疫分析（SS-RTP-IA）新方法奠定了相应的实验基础。

Study on the Properties of Solid Substrate Room Temperature Phosphorescence of the Antibody Labeled with Fluorescein Isothiocynate

The properties of room temperature phosphorescence (RTP) of the labeled goat anti-human antibody(with fluorescein isothiocynate GAHAb-FITC) and labeled rabit anti-goat antibody(RAGAb-FITC) with FITC in different dilution ratios on many kinds of solid substrates were studied in detail. It is found that on the polyamide membrane, only using Pb(Ac)2 as a heavy atom perturber, both the labeled antibodies and their antigen-antibody conjugated compound with the human immunoglobulin G remained excellent RTP properties of isothiocyanate fluorescein, lambda ex(max)/lambda em(max) = 525/650 nm, the relation of RTP intensity to the concentration was linear in a certain range. The conditions for immunological reaction and RTP emission were optimized. The result means that it is possible to develop a new immunoassay named as solid substrate room temperature phosphorescence immunoassay (SS-RTP-IA), which combines both the specificity of immunological reaction and high sensitivity of luminescence analysis.