Multiplex immunoassay for in vitro characterization of acellular pertussis antigens in combination vaccines
Multiplex immunoassay for in vitro characterization of acellular pertussis antigens in combination vaccines
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DOI：
10.1016/j.vaccine.2016.01.012
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发表时间：
2016-02-17
期刊：
影响因子：
5.5
通讯作者：
Baudner, Barbara C.
中科院分区：
文献类型：
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作者：
Agnolon, Valentina;Bruno, Cristina;Baudner, Barbara C.
为了确保抗原和佐剂的物理、化学和生物完整性，需要对疫苗进行表征。目前的分析方法大多需要从铝基佐剂中完全解吸抗原，并且并不总是适合于区分多价制剂中的单个抗原。在这里，Luminex技术被提出用于改进疫苗特性的分析。为验证概念，选择以氢氧化铝为佐剂的TdaP（破伤风、白喉和无细胞百日咳）组合作为模型配方，同时定量测定无细胞百日咳（aP）抗原在佐剂表面的吸附水平。该试验使用特异性抗体与磁微球结合，为每种百日咳抗原提供独特的数字签名，允许同时识别整个疫苗中的各自抗原，避免了费力的佐剂分离程序。在0.78-50 ng/mL范围内实现了TdaP疫苗中aP抗原的准确和可重复性定量，同时提供了抗原的特性、数量和对氢氧化铝的吸附程度的信息。目前的研究可以进一步被视为建立体外效价测定的模型，从而支持根据3Rs概念取代动物试验。(C) 2016 Elsevier Ltd.版权所有。
Vaccines characterization is required to ensure physical, chemical, and biological integrity of antigens and adjuvants. Current analytical methods mostly require complete antigen desorption from aluminum-based adjuvants and are not always suitable to distinguish individual antigens in multivalent formulations. Here, Luminex technology is proposed to improve the analytics of vaccine characterization. As proof of concept, TdaP (tetanus, diphtheria and acellular pertussis) combination, adjuvanted with aluminum hydroxide, was chosen as model formulation to quantify and determine the level of adsorption of acellular pertussis (aP) antigens onto adjuvant surface at the same time. The assay used specific antibodies bound to magnetic microspheres presenting unique digital signatures for each pertussis antigen, allowing the simultaneous recognition of respective antigens in the whole vaccine, avoiding laborious procedures for adjuvant separation. Accurate and reproducible quantification of aP antigens in TdaP vaccine has been achieved in the range 0.78-50 ng/mL, providing simultaneously information on antigen identity, quantity, and degree of adsorption to aluminum hydroxide. The current study could further be considered as a model to set up in vitro potency assays thus supporting the replacement of animal tests accordingly to the 3Rs concept. (C) 2016 Elsevier Ltd. All rights reserved.