The investigation for high sensitive chemi luminescence method using nanoreactor as a marker

制备了包封辣根过氧化物酶（HRP）的脂质体，将脂质体内水相应用于纳米反应器，进行HRP催化化学发光（CL）反应。磷脂酰胆碱（PC）、磷脂酰甘油二肉豆蔻酰基（DMPG）和胆固醇作为脂质体的组成部分。PC: DMPG: Chel的摩尔比为8:1:1。采用聚碳酸酯滤池挤压法制备酶陷脂质体。采用均质酸γ-内酯（HAL）和鲁米诺作为CL试剂，测定脂质体中辣根过氧化物酶（HRP）的含量。用Triton X-100裂解HRP捕获的脂质体后检测HRP。对HAL CL中HRP的反应率、检出限和线性范围进行了比较。与ρ-IP增强CL相比，HAL CL的最大光发射出现得更快，从而显著缩短了CL的测量时间。HRP在HAL CL中的检出限与在ρ-IP增强甲醇CL中的检出限相同。与ρ-IP增强鲁米诺CL相比，HAL CL中HRP的校准曲线线性范围提高了50倍。结果表明，在测定脂质体中HRP含量时，HAL CL优于ρ-IP增强鲁米诺CL。

Liposomes encapsulated horseradish peroxidase(HRP) were prepared for applying the inner water phase in liposome to the nanoreactor for HRP-catalyzed chemiluminecence(CL) reaction. Phosphatidylcholine(PC), phosphatidylglycerol dimyristoyl(DMPG) and cholesterol were used as a component for liposome. The mole % of PC : DMPG : Chel was 8:1:1. The HRP-trapped liposome was prepared by extrusion method with polycarbonate filter. Homogentisic acid γ-lactone(HAL) and luminol were used as a CL reagent for the determination of horseradish peroxidase (HRP) encapsulated in liposomes. HRP was detected after the lysis of HRP-trapped liposomes with Triton X-100. CL response rate, detection limit and linear range of calibration curve for HRP in HAL CL were compared with those in ρ-iodophenol (ρ-IP) enhanced luminol CL. Maximal light emission in HAL CL appeared more rapidly compared to that in ρ-IP enhanced CL methods, thus resulting in remarkable reduction of CL measurement time. The detection limit for HRP in HAL CL was the same as that in ρ-IP enhanced lnminol CL. The linear range of calibration curve for HRP in HAL CL was improved by factors of 50 compared with that in ρ-IP enhanced luminol CL. From these results, it was found that HAL CL were superior to ρ-IP enhanced luminol CL for the determination of HRP encapsulated in liposomes.