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

制备了包裹辣根过氧化物酶(HRP)的脂质体，将脂质体中的内水相应用于纳米反应器中，用于HRP催化的化学发光(CL)反应。以磷脂酰胆碱(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成分。PC：DMPG：CHEL的摩尔比为8：1：1。以均一酸、γ-内酯和鲁米诺为化学发光试剂，测定了脂质体中辣根过氧化物酶的含量。用Triton X-100溶解HRP脂质体后，检测HRP。比较了HAL CL和ρ-碘苯酚(ρ-IP)增敏鲁米诺CL中HRP的CL响应率、检出限和标准曲线的线性范围。与ρ-IP增强化学发光方法相比，HAL化学发光的最大发光速度更快，从而显著缩短了化学发光测量时间。HAL CL与ρ-IP增强型Inminol CL的检出限相同。与ρ-IP增强型鲁米诺CL相比，HAL CL中HRP的线性范围提高了50倍。结果表明，HAL CL法优于ρ-IP增强型鲁米诺CL法测定脂质体中的辣根过氧化物酶。

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.