Grier Thomas J, Hall Dawn M, Duncan Elizabeth A, Coyne Terrance C
Research and Development Laboratory, Greer Laboratories, Inc, Lenoir, North Carolina.
Research and Development Laboratory, Greer Laboratories, Inc, Lenoir, North Carolina.
Ann Allergy Asthma Immunol. 2015 Mar;114(3):233-9. doi: 10.1016/j.anai.2014.11.021. Epub 2015 Jan 9.
Recent studies have shown that Alternaria and German cockroach allergens can be degraded by endogenous proteases from other insect and fungal extracts when combined for immunotherapy, but data supporting the compatibilities of other high-protease products in comparable mixtures have not been reported.
To assess the stabilities and compatibilities of Aspergillus fumigatus and American cockroach allergens after mixing with protease-rich extracts from other insects or fungi at concentrations similar to those recommended for subcutaneous immunotherapy.
Mixtures containing A fumigatus, American cockroach, and other fungal or insect extracts were evaluated by quantitative (enzyme-linked immunosorbent assays) and qualitative (immunoblotting) methods. Test mixtures and control samples at 10% to 50% glycerin concentrations were analyzed after storage for up to 12 months at 2°C to 8°C.
Moderate to high recoveries of Aspergillus extract activities were retained in control samples and extract mixtures under all conditions examined. American cockroach extract controls were partly degraded at 10% to 25% glycerin, and cockroach allergen compatibilities were decreased significantly in mixtures with several fungal extracts at 25% glycerin. Mixing with other insects did not compromise the stability of American cockroach allergens at 25% to 50% glycerin.
Aspergillus extracts exhibited favorable stabilities after mixing with other high-protease products. American cockroach extract potencies were unstable in less than 50% glycerin, even in the absence of other protease-containing allergens, and were destabilized in mixtures with several fungal extracts. Addition of fungal and insect extracts to separate treatment vials or preparation of fungal-insect mixtures at elevated glycerin concentrations might be necessary to produce compatible patient formulations for allergen immunotherapy injections.
最近的研究表明,链格孢属真菌和德国小蠊过敏原在联合用于免疫治疗时可被其他昆虫和真菌提取物中的内源性蛋白酶降解,但尚未有数据支持其他高蛋白酶产品在类似混合物中的兼容性。
评估烟曲霉和美洲大蠊过敏原与其他昆虫或真菌富含蛋白酶的提取物以皮下免疫治疗推荐浓度混合后的稳定性和兼容性。
通过定量(酶联免疫吸附测定)和定性(免疫印迹)方法评估含有烟曲霉、美洲大蠊以及其他真菌或昆虫提取物的混合物。在2℃至8℃储存长达12个月后,分析甘油浓度为10%至50%的测试混合物和对照样品。
在所有检测条件下,对照样品和提取物混合物中烟曲霉提取物活性的回收率保持在中等至高水平。美洲大蠊提取物对照在甘油浓度为10%至25%时部分降解,在甘油浓度为25%时,与几种真菌提取物的混合物中美洲大蠊过敏原的兼容性显著降低。与其他昆虫混合不会影响美洲大蠊过敏原在甘油浓度为25%至50%时的稳定性。
烟曲霉提取物与其他高蛋白酶产品混合后表现出良好的稳定性。美洲大蠊提取物在甘油浓度低于50%时效力不稳定,即使不存在其他含蛋白酶的过敏原,并且在与几种真菌提取物的混合物中会失稳。可能需要将真菌和昆虫提取物添加到单独的治疗小瓶中,或者在较高甘油浓度下制备真菌 - 昆虫混合物,以生产用于过敏原免疫治疗注射的兼容患者制剂。