人用疫苗氢氧化铝佐剂吸附力的质量控制初探

目的 建立氢氧化铝佐剂吸附力的相关质控方法.方法 对3批国产氢氧化铝佐剂与丹麦铝佐剂（Alhydrogel）同时进行粒径、比表面积、零电荷点的测定,并比较两种佐剂对不同浓度牛血清白蛋白（bovine serum albumin,BSA）及百日咳丝状血凝素（filamentous hemagglutinin,FHA）的吸附力.结果 测定显示,3批国产铝佐剂50％体积百分比（DV50）的粒径分布分别为4.49、5.83和6.28 μm;比表面积分别为1 602、1 093和1 014 m^2/kg;零电荷点分别为10.2、10.3和10.2.丹麦铝佐剂DV50的粒径为4.58 μm,比表面积为2 230 m^2/kg,零电荷点为8.5.3批国产和1批丹麦铝佐剂（5 g/L）对2 g/L BSA的吸附率分别为98.69％、99.41％、99.72％和98.04％,均符合欧洲药典的吸附力合格标准.国产和丹麦铝佐剂（1.3 g/L）对50～1 000 mg/L FHA的吸附率均在97％以上.结论 根据对国产氢氧化铝佐剂相关质量指标的研究,初步建立了氢氧化铝佐剂吸附力的质控标准.     

对氢氧化铝和磷酸钙作为疫苗佐剂时的毒性研究

作者配制了3mg/ml的氢氧化铝凝胶(Al-gel)、氢氧化铝悬液(Al—SUS)、磷酸钙凝胶(Ca-gel)和磷酸钙悬液(Ca-SUS)作为试验材料。通过测定血管通透性来评价它们引起水肿的程度,即在豚鼠静脉注射含1％伊文斯蓝的生理盐水后立即皮内注射试验材料,5小时后从皮内切除注射部位的伊文斯蓝块,在620nm下测定光吸收值,以表示血管通透性强度。     

喷雾式添加氨水对氢氧化铝佐剂质量一致性的影响

目的  通过喷雾式添加氨水制备氢氧化铝佐剂,分析其对佐剂质量批间一致性的影响。 方法  分别以喷雾和流加方式添加氨水至三氯化铝溶液各制备3批氢氧化铝佐剂,比较两种氨水添加工艺制备的佐剂粒径、pH、外观等质量指标。 结果  喷雾方式添加氨水制备的3批佐剂平均粒径为（96.7±8.6） nm,pH为4.93±0.02,透析后沉淀量为（20.9±0.4） g/L;流加方式添加氨水制备的3批佐剂平均粒径为（161.0±24.7） nm,pH为4.85±0.06,透析后沉淀量为（105.6±13.8） g/L。相较于流加,以喷雾式添加氨水制备的3批佐剂为均一的乳白色悬液,粒径更小,透析后透析袋底部大颗粒沉淀明显减少,批间一致性更好。 结论  采用喷雾方式将氨水加入三氯化铝溶液中,可使氨水与三氯化铝溶液接触更加充分,产物更加均匀,有助于提高氢氧化铝佐剂外观、粒径、pH等质量指标的批间一致性。     

疫苗佐剂研究进展

在疫苗生产过程中,使用适当的佐剂,不再是单纯地提高机体对抗原的免疫应答水平,而更着重于选择性地诱导产生有效防御相应病原体感染的特异性应答的类型。本文从辅助性T(Th)淋巴细胞亚型及特异性免疫应答类型角度,对新近研究开发的重要选择性佐剂(油性乳剂、脂多糖、皂苷、脂质体以及细胞因子)作一介绍。     

疫苗佐剂研究进展

在疫苗生产过程中，使用适当的佐剂，不再是单纯地提高机体对抗原的免疫应答水平，而更着重于选择性地诱导产生有效防御相应病原体感染的特异性应答的类型。本文从辅助性Ｔ（Ｔｈ）淋巴细胞亚型及特异性免疫应答类型角度，对新近研究开发的重要选择性佐剂（油性乳剂、脂多糖、皂苷、脂质体以及细胞因子）作一介绍。     

不可忽视的疫苗佐剂

尽管国产乙型脑炎减毒活疫苗通过了WHO预认证,也意味着国产疫苗的安全性又向前迈进了一步,但是这一消息并未能吹散笼罩在人们头上那团“疫苗安全性问题”的乌云。媒体对疫苗后遗症的系列报道的后遗效应还在继续刺激公众乃至业内人士疼痛的神经。     

疫苗佐剂最新研究进展

疫苗佐剂能够提高机体对抗原的适应性免疫应答,在疫苗的研发中具有重要的作用。随着药物研发的不断进步,近年来国内外出现了许多新型佐剂。本文分类阐述了近年来疫苗佐剂的最新研究进展及应用概况,总结了佐剂对免疫系统的影响和作用机制、临床有效性和不良反应,以及新型佐剂研发最新关注热点。在寻找并设计理想佐剂的同时我们不能忽略安全问题,应深入研究佐剂的作用机制及评价模型,全面评估其安全性。     

脂质体佐剂疫苗研究进展

脂质体作为蛋白质及多肽抗原的佐剂和载体已被广泛应用。通过综述脂质体在细菌、病毒、寄生虫和肿瘤疫苗中的研究应用 ,提示脂质体在疫苗的研制中具有广阔的应用前景     

疫苗佐剂的研究进展

新一代疫苗，尤其是重组蛋白或DNA，比传统疫苗反应性弱，免疫原性也小。因此，迫切需要开展和改进疫苗佐剂。根据佐剂的主要作用机制可以将其分为两大类：疫苗输送系统及免疫刺激性佐剂。疫苗输送系统通常为微粒，功能主要是使相关抗原进入抗原提呈细胞。免疫刺激佐剂主要由病原体衍生，模拟病原体相关分子模式，激活先天免疫系统的细胞。更多有效的佐剂的发现促进了对于癌症和慢性感染性疾病的预防和治疗性疫苗的发展。此外，新佐剂的发展也使疫苗经粘膜应用成为可能。     

脂质体佐剂疫苗研究进展

脂质体作为蛋白质及多肽抗原的佐剂和载体已被广泛应用。通过综述脂质体在细菌、病毒、寄生虫和肿瘤疫苗中的研究应用，提示脂质体在疫苗的研制中具有广阔的应用前景。     

Measuring the surface area of aluminum hydroxide adjuvant

The traditional method of determining surface area, nitrogen gas sorption, requires complete drying of the sample prior to analysis. This technique is not suitable for aluminum hydroxide adjuvant because it is composed of submicron, fibrous particles that agglomerate irreversibly upon complete removal of water. In this study, the surface area of a commercial aluminum hydroxide adjuvant was determined by a gravimetric/FTIR method that measures the water adsorption capacity. This technique does not require complete drying of the adjuvant. Five replicate determinations gave a mean surface area of 514 m(2)/g and a 95% confidence interval of 36 m(2)/g for a commercial aluminum hydroxide adjuvant. The X-ray diffraction pattern and the Scherrer equation were used to calculate the dimensions of the primary crystallites. The average calculated dimensions were 4.5 x 2.2 x 10 nm. Based on these dimensions, the mean calculated surface area of the commercial aluminum hydroxide adjuvant was 509 m(2)/g, and the 95% confidential interval was 30 m(2)/g. The close agreement between the two surface area values indicates that either method may be used to determine the surface area of aluminum hydroxide adjuvant. The high surface area, which was determined by two methods, is an important property of aluminum hydroxide adjuvants, and is the basis for the intrinsically high protein adsorption capacity.     

Inhibition of aggregation of aluminum hydroxide adjuvant during freezing and drying

Aluminum-salt adjuvants are widely used to increase immunogenicity of recombinant protein vaccines. However, when vaccines formulated with these adjuvants are frozen or lyophilized, losses of efficacy are often reported. This loss of potency is usually attributed to the aggregation of adjuvant particles during processing. In this study, we examine the aggregation behavior of Alhydrogel, a commercial aluminum hydroxide adjuvant, during freeze-thawing and freeze-drying. By cooling Alhydrogel formulations at faster rates or by the addition of sufficient amounts of a glass forming excipient such as trehalose, aggregation of Alhydrogel, can be prevented or minimized. We propose that freeze-concentration of buffer salts induces modifications in adjuvant surface chemistry and crystallinity, which in turn favor aggregation. These modifications, and the resulting aggregation of Alhydrogel particles can be minimized through choice of buffer ions, or kinetically inhibited by rapidly forming a glassy state during freezing.     

氢氧化钠质量标准中钾盐、铝盐检查项的探讨与研究

目的:探讨《中国药典》2020年版四部氢氧化钠标准中钾盐、铝盐检查项目的合理性和修订意见。方法:寻找灵敏度更高、专属性更强和操作更简便的方法,对钾盐和铝盐进行检查。结果:四苯硼钠方法检查钾盐,灵敏度更高。采用专属性更强的铝试剂检验方法,替代炽灼残渣方法检测铝盐。结论:本文建立的方法能对钾盐和铝盐进行更灵敏,专属性更强的检验,操作简便。     

Influence of protein conformation and adjuvant aggregation on the effectiveness of aluminum hydroxide adjuvant in a model alkaline phosphatase vaccine

The mechanism(s) of the enhancement of the immune response by addition of aluminum salt adjuvants to parenterally administered protein-based vaccines is still the subject of debate. It has been hypothesized, however, that destabilization of the antigen structure on the surface of the adjuvant may be important for eliciting immune response. Also, it has been suggested that immune response to adjuvanted vaccines is reduced if the adjuvant particles become aggregated before administration because of processing steps such as freeze-drying. In this study, we tested these hypotheses and examined the immune response in a murine model to various liquid, freeze-dried, and spray freeze-dried formulations of a model vaccine, bovine intestinal alkaline phosphatase adsorbed on aluminum hydroxide. Enzymatic activity of the alkaline phosphatase was used as a sensitive indicator of intact native antigen structure. By manipulating the secondary drying temperature during lyophilization, vaccines were produced with varying levels of alkaline phosphatase enzymatic activity and varying degrees of adjuvant aggregation, as assessed by particle size distribution. Anti-alkaline phosphatase titers observed in immunized mice were independent of both the antigen's retained enzymatic activity and the vaccine formulation's mean particle diameter.     

病毒性疫苗渗透压摩尔浓度的质量控制

目的  通过检测6种病毒性疫苗成品的渗透压摩尔浓度,比较不同疫苗检测均值的差异,并观察同种疫苗检测值的批间稳定性,为增加病毒性疫苗质量控制手段提供依据。方法  采用冰点下降法检测麻疹减毒活疫苗、风疹减毒活疫苗、麻疹腮腺炎联合减毒活疫苗、麻疹腮腺炎风疹联合减毒活疫苗、水痘减毒活疫苗、流感病毒裂解疫苗的渗透压摩尔浓度,对检测值进行统计学处理,计算变异系数。以麻疹腮腺炎风疹联合减毒活疫苗的渗透压摩尔浓度检测均值作为对照,进行方差齐性检验及假设检验,比较各疫苗检测均值的差异。结果  麻疹腮腺炎联合减毒活疫苗与对照相比,均值差异无统计学意义（t=1.66,P>0.05）;麻疹减毒活疫苗、风疹减毒活疫苗、水痘减毒活疫苗及流感病毒裂解疫苗与对照相比,均值差异均有统计学意义（Z>1.96,P<0.001）。同种疫苗批间渗透压摩尔浓度较为稳定,变异系数均<3%,变化幅度能控制在90%~110%均值范围内。结论  6种病毒性疫苗渗透压摩尔浓度存在一定差异,但同种疫苗检测值批间稳定性较好,因此,应根据不同疫苗的渗透压摩尔浓度,分别制定质量控制标准。     

Effect of the degree of phosphate substitution in aluminum hydroxide adjuvant on the adsorption of phosphorylated proteins

Aluminum hydroxide adjuvant was pretreated with six concentrations of potassium dihydrogen phosphate to produce a series of adjuvants with various degrees of phosphate substitution for surface hydroxyl. The adsorption of three phosphorylated proteins (alpha casein, dephosphorylated alpha casein, and ovalbumin) by the phosphate-treated aluminum hydroxide adjuvants was studied. The phosphorylated proteins were adsorbed by ligand exchange of phosphate for hydroxyl even when an electrostatic repulsive force was present. However, the extent (adsorptive capacity) and strength (adsorptive coefficient) of adsorption was inversely related to the degree of phosphate substitution of the aluminum hydroxide adjuvant. Exposure of vaccines containing aluminum hydroxide adjuvant and phosphorylated antigens to phosphate ion in the formulation or during manufacture should be minimized to produce maximum adsorption of the antigen.