Recombinant HA1 produced in E. coli forms functional oligomers and generates strain-specific SRID potency antibodies for pandemic influenza vaccines
Recombinant HA1 produced in E. coli forms functional oligomers and generates strain-specific SRID potency antibodies for pandemic influenza vaccines
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DOI：
10.1016/j.vaccine.2011.06.014
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发表时间：
2011-08-05
期刊：
影响因子：
5.5
通讯作者：
Golding, Hana
中科院分区：
文献类型：
--
作者：
Khurana, Surender;Larkin, Christopher;Golding, Hana
疫苗生产和启动大规模疫苗接种是快速应对新型流感大流行的关键因素。在2009-2010年H1N1流感大流行期间，发现了几个瓶颈，包括疫苗效力试剂的延迟供应。目前，用于单向免疫扩散（SRID）效价测定的抗血清是在用菠萝蛋白酶处理鸡蛋中生长的流感病毒后释放和纯化的HA反复免疫的绵羊中产生的。这种方法是2009年大流行性H1N1（H1N1 pdm 09）效价试剂开发的主要瓶颈。需要替代方法来制备HA免疫原，以在尽可能短的时间内产生SRID试剂。在这项研究中，我们发现，正确折叠的重组HA 1球状结构域（rHA 1）从几个A型病毒，包括H1N1 pdm 09和两个H5 N1病毒可以有效地使用细菌表达系统和随后的纯化。rHA 1蛋白显示形成三聚体的功能性寡聚体，类似于病毒衍生的HA，并在兔和绵羊中引起高滴度的中和抗体。重要的是，免疫血清在SRID测定中以剂量依赖性方式与参考抗原形成沉淀环。用rHA 1产生的羊血清测定的来自不同制造商（和几个批次）的多种H1N1疫苗产品中的HA含量与用NIBSC传统产生的羊血清获得的值相似。我们的结论是，细菌表达的重组HA 1蛋白可以快速生产，并用于产生SRID效价试剂后不久，新的流感病毒株与大流行的潜力被确定。爱思唯尔有限公司出版
Vaccine production and initiation of mass vaccination is a key factor in rapid response to new influenza pandemic. During the 2009-2010 H1N1 pandemic, several bottlenecks were identified, including the delayed availability of vaccine potency reagents. Currently, antisera for the single-radial immunodiffusion (SRID) potency assay are generated in sheep immunized repeatedly with HA released and purified after bromelain-treatment of influenza virus grown in eggs. This approach was a major bottleneck for pandemic H1N1 (H1N1pdm09) potency reagent development in 2009. Alternative approaches are needed to make HA immunogens for generation of SRID reagents in the shortest possible time. In this study, we found that properly folded recombinant HA1 globular domain (rHA1) from several type A viruses including H1N1pdm09 and two H5N1 viruses could be produced efficiently using a bacterial expression system and subsequent purification. The rHA1 proteins were shown to form functional oligomers of trimers, similar to virus derived HA, and elicited high titer of neutralizing antibodies in rabbits and sheep. Importantly, the immune sera formed precipitation rings with reference antigens in the SRID assay in a dose-dependent manner. The HA contents in multiple H1N1 vaccine products from different manufacturers (and in several lots) as determined with the rHA1-generated sheep sera were similar to the values obtained with a traditionally generated sheep serum from NIBSC. We conclude that bacterially expressed recombinant HA1 proteins can be produced rapidly and used to generate SRID potency reagents shortly after new influenza strains with pandemic potential are identified. Published by Elsevier Ltd.