氢氧化铝佐剂吸附肠道病毒71型灭活病毒的特性  

Absorption characteristics of aluminum hydroxide adjuvant with inactivated enterovirus 71

作　　者：严皎 赵勇 贺凌煜 納锐雄 李亚东 袁明翠 蒋曦 易力 YAN Jiao;ZHAO Yong;HE Lingyu;NA Ruixiong;LI Yadong;YUAN Mingcui;JIANG Xi;YI Li(Institute of Medical Biology,Chinese Academy of Medical Sciences,Kunming 650000,Yunnan Province,China)

机构地区：[1]中国医学科学院医学生物学研究所、质量检定室国家药品监督管理局疫苗及生物制品质量控制与评价重点实验室,云南昆明650000

出　　处：《中国生物制品学杂志》2023年第6期651-656,共6页Chinese Journal of Biologicals

基　　金：云南省重大科技专项(202002AA100009)。

摘　　要：目的探讨氢氧化铝佐剂吸附肠道病毒71型(enterovirus 71,EV71)灭活病毒的特性。方法采用透射电镜、凝胶排阻层析高效液相色谱(size exclusion chromatography HPLC,SEC-HPLC)及动态光散射(dynamic light scatter,DLS)技术分析灭活EV71颗粒的形态、纯度及粒径分布,并以透射电镜观察氢氧化铝佐剂纳米颗粒形态。以灭活EV71抗原含量(4000、6000、8000、10000、11000、12000、13000、14000、20000、30000 U/mL)、氢氧化铝佐剂浓度[0.35、0.25、0.17、0.085 mg/mL(按铝含量计)]、吸附时间(0、30及120 min)、离子强度(NaCl浓度0.15、0.75和1.25 mol/L)及磷铝摩尔比(P/Al,0.15、0.64、2.08及7.87)为变量,考察氢氧化铝佐剂吸附灭活EV71抗原后所呈现的吸附特性。结果灭活EV71颗粒主要以完整的病毒颗粒形式存在,直径约30 nm;氢氧化铝佐剂具有纳米化的特征,粒径分布在200~700 nm之间。≤11000 U/mL浓度的灭活EV71抗原可被0.35 mg/mL(按铝含量计)的氢氧化铝佐剂完全吸附,上清中未检测到游离抗原,从12000 U/mL起,随抗原量的增加,上清中游离抗原含量也增加;各浓度氢氧化铝佐剂能完全吸附250 U/mL的灭活EV71抗原,上清中未检测到游离抗原,不同孵育时间的吸附效果一致。在单剂量疫苗氢氧化铝含量(0.35 mg/mL,按铝含量计)及灭活EV71抗原含量(250 U/mL)条件下,NaCl离子强度对灭活EV71吸附无影响,而磷酸离子浓度明显影响吸附。结论单剂量疫苗中的氢氧化铝佐剂可完全吸附灭活EV71抗原,基团置换在其中发挥了重要作用。Objective To explore the absorption characteristics of aluminum hydroxide adjuvant with inactivated enterovirus 71(EV71).Methods The morphology,purity and particle size distribution of inactivated EV71 particles were analyzed by transmission electron microscope,size exclusion chromatography HPLC(SEC-HPLC)and dynamic light scatter(DLS),and the morphology of aluminum hydroxide adjuvant nanoparticles was observed by transmission electron microscope.Using inactivated EV71 antigen content(4000,6000,8000,10000,11000,12000,13000,14000,20000,30000 U/mL),aluminum hydroxide adjuvant concentration[0.35,0.25,0.17,0.085 mg/mL(aluminum content)],adsorption time(0,30 and 120 min),ionic strength(sodium chloride concentration of 0.15,0.75 and 1.25 mol/L)and phosphorus-aluminum molar ratio(P/Al,0.15,0.64,2.08 and 7.87)as variables,the adsorption characteristics of aluminum hydroxide adjuvant with inactivated EV71 antigen were investigated.Results Inactivated EV71 particles mainly existed in the form of intact virus particles with a diameter of about 30 nm;Aluminum hydroxide adjuvant showed the characteristics of nanocrystallization,and the particle size was distributed within 200~700 nm.The inactivated EV71 antigen at the concentration of no more than 11000 U/mL was completely absorbed by aluminum hydroxide adjuvant of 0.35 mg/mL(aluminum content),and no free antigen was detected in the supernatant;while from 12000 U/mL,the content of free antigen in the supernatant increased with the increase of antigen content;the inactivated EV71 antigen of 250 U/mL was completely absorbed by various concentrations of aluminum hydroxide adjuvant,and no free antigen was detected in the supernatant;the adsorption effect of aluminum hydroxide adjuvant consistent after incubation for different time,and no free antigen was detected in the supernatant.Under the conditions of single dose of vaccine with aluminum hydroxide content(0.35 mg/mL)and inactivated EV71 antigen content(250 U/mL),sodium chloride ion strength had no effect on the adsorption of i

关 键 词：氢氧化铝佐剂 肠道病毒71型 吸附 灭活病毒 

分 类 号：R392-33[医药卫生—免疫学] R373.2[医药卫生—基础医学]