Martorell D, Siebert S T, Durst R A
Analytical Chemistry Laboratories, Cornell University, Geneva, New York 14456-0462, USA.
Anal Biochem. 1999 Jul 1;271(2):177-85. doi: 10.1006/abio.1999.4134.
The feasibility of utilizing dehydrated liposomes in the development of a simple immunoassay device for point-of-care diagnostics or field assays was demonstrated. The recovery of liposomes after a cycle of dehydration and rehydration was studied using biotin-tagged, dye-loaded liposomes with antibiotin antibodies immobilized in a defined zone on nitrocellulose strips. Liposomes were vacuum-dehydrated on the strip at a location below the antibiotin zone. The strip was placed in a tube containing a carrier solution and capillary action brought the solution to the dehydrated liposomes, rehydrated them, and caused them to migrate to the antibody zone where intact liposomes were captured and measured optically. High concentrations of either trehalose or sucrose external to the liposomes and both polyvinylpyrrolidone and gelatin in the membrane blocking reagent were essential for preservation of the dehydrated/rehydrated liposomes on nitrocellulose. Between 70 and 80% of the liposomes were recovered on the nitrocellulose strips after a cycle of dehydration and rehydration. The dehydrated liposomes on the strips were stable for at least 1 year when stored in vacuum-sealed plastic bags at 4 degrees C. The technique was successfully applied to the development of a rapid one-step strip immunoassay for biotin.
已证明利用脱水脂质体开发用于即时诊断或现场检测的简单免疫分析装置的可行性。使用生物素标记、负载染料且抗生物素抗体固定在硝酸纤维素条特定区域的脂质体，研究了脱水和再水化循环后脂质体的回收率。脂质体在抗生物素区域下方的条带上进行真空脱水。将条带置于含有载体溶液的试管中，毛细管作用使溶液到达脱水脂质体，使其再水化，并使其迁移至抗体区域，在那里完整的脂质体被捕获并进行光学测量。脂质体外高浓度的海藻糖或蔗糖以及膜封闭试剂中的聚乙烯吡咯烷酮和明胶对于在硝酸纤维素上保存脱水/再水化脂质体至关重要。经过脱水和再水化循环后，70%至80%的脂质体在硝酸纤维素条带上得以回收。条带上的脱水脂质体在4℃真空密封塑料袋中储存时至少稳定1年。该技术已成功应用于生物素快速一步条带免疫分析的开发。
