Jack Jumper Allergy Program, Royal Hobart Hospital, Hobart, Tas., Australia.
Department of Pharmacy, Royal Hobart Hospital, Hobart, Tas., Australia.
Clin Exp Allergy. 2017 Nov;47(11):1478-1490. doi: 10.1111/cea.12987. Epub 2017 Aug 31.
Allergen immunotherapy uses pharmaceutical preparations derived from naturally occurring source materials, which contain water-soluble allergenic components responsible for allergic reactions. The success of in vivo and in vitro diagnoses in allergen sensitization and allergen immunotherapy largely depends on the quality, composition and uniformity of allergenic materials used to produce the active ingredients, and the formulation employed to prepare finished products.
We aimed to examine the factors influencing batch-to-batch consistency of Jack Jumper (Myrmecia pilosula) ant venom (JJAV) in the form of active pharmaceutical ingredient (AI) and informed whether factors such as temperature, artificial light and container materials influence the quality of JJAV AIs. We also aimed to establish handling and storage requirements of JJAV AIs to ensure preservation of allergenic activities during usage in the diagnosis of allergen sensitization and in allergen immunotherapy.
The quality and consistency of JJAV AIs were analysed using a combination of bicinchoninic acid assay for total protein quantification, HPLC-UV for JJAV allergen peptides quantification, ELISA inhibition for total allergenic potency, SDS-PAGE, AU-PAGE and immunoblot for qualitative assessment of JJAV components, and Limulus Amebocyte Lysate assay for the quantification of endotoxin concentration. API-ZYM and Zymogram assays were used to probe the presence of enzymatic activities in JJAV.
Pharmaceutical-grade JJAV for allergen immunotherapy has good batch-to-batch consistency. Temporary storage at 4°C and light exposure do not affect the quality of JJAV. Exposure to temperature above 40°C degrades high MW allergens in JJAV. Vials containing JJAV must be stored frozen and in upright position during long-term storage.
We have identified factors, which can influence the quality and consistency of JJAV AIs, and provided a framework for appropriate handling, transporting and storage of JJAV to be used for the diagnosis of allergen sensitization and in AIT.
变应原免疫治疗使用来源于天然来源材料的药物制剂，其中包含负责过敏反应的水溶性变应原成分。变应原致敏和变应原免疫治疗的体内和体外诊断的成功在很大程度上取决于用于生产活性成分的变应原材料的质量、组成和均一性，以及用于制备成品的配方。
我们旨在研究影响跳蚁（Myrmecia pilosula）毒液（JJAV）作为活性药物成分（AI）的批次间一致性的因素，并确定温度、人工光和容器材料等因素是否会影响 JJAV AI 的质量。我们还旨在确定 JJAV AI 的处理和储存要求，以确保在用于诊断变应原致敏和变应原免疫治疗的过程中保持变应原活性。
使用组合的二辛可宁酸测定法（bicinchoninic acid assay）进行总蛋白定量、高效液相色谱-紫外分光光度法（HPLC-UV）进行 JJAV 变应原肽定量、酶联免疫吸附抑制试验（ELISA inhibition）进行总变应原效力、SDS-PAGE、AU-PAGE 和免疫印迹进行 JJAV 成分的定性评估，以及鲎变形细胞溶解物试验（Limulus Amebocyte Lysate assay）进行内毒素浓度定量。API-ZYM 和 Zymogram 测定法用于探测 JJAV 中酶活性的存在。
用于变应原免疫治疗的制药级 JJAV 具有良好的批次间一致性。4°C 下的临时储存和光照暴露不会影响 JJAV 的质量。暴露于 40°C 以上的温度会降解 JJAV 中的高分子量变应原。含有 JJAV 的小瓶在长期储存期间必须冷冻储存并保持垂直位置。
我们已经确定了会影响 JJAV AI 质量和一致性的因素，并为 JJAV 的适当处理、运输和储存提供了框架，以用于变应原致敏的诊断和 AIT。
