藻红蛋白标记抗鸡IgG荧光抗体的高效制备

Preparation of phycoerythrin-labelled anti-avian-IgG-antibody

藻红蛋白的斯托克位移大,荧光量子产率高,荧光明亮,是一种应用前景广阔的荧光染料。制约藻红蛋白应用的主要因素是分离纯化困难及其成品售价昂贵。通过我们已经建立的藻红蛋白低成本高效制备平台,使其应用于预防兽医学检测成为可能。首次采用适宜摩尔浓度的交联剂SPDP将藻红蛋白与抗鸡IgG抗抗体以摩尔比1∶1高效交联,经高效液相色谱法纯化制备出藻红蛋白标记的抗鸡IgG荧光抗抗体。本研究对制备的荧光抗抗体从荧光特性、吸收光谱特性、抗体活性、纯度及稳定性等方面进行了鉴定。结果表明,藻红蛋白标记的抗鸡IgG荧光抗体为电泳纯,荧光明亮,抗体活性高,性质稳定,可用于鸡传染病的间接免疫荧光检测。

Phycoerythrin is an excellent fluorescent dye because of its high fluorescence quantum yield and large stokes shift. However, complicated purification procedure and high price of phycoerythrin hindered its wide application. In this study, an efficient method for preparation of R-phycoerythrin-labelled anti-avian IgG autibody was established using optimum concentration of crosslinker SPDP. The labelled antibody was purified with HPLC and tested for absorption and fluorescence spectra, bioreactivity, and stability. The result showed that the purified labelled antibody had good bioreactivity and stability, which could be widely applied for immunofluorescence detection of avian infectious diseases.