Formulation development and comparability studies with an aluminum-salt adjuvanted SARS-CoV-2 Spike ferritin nanoparticle vaccine antigen produced from two different cell lines.
Formulation development and comparability studies with an aluminum-salt adjuvanted SARS-CoV-2 Spike ferritin nanoparticle vaccine antigen produced from two different cell lines.
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使用由两种不同细胞系生产的铝盐佐剂 SARS-CoV-2 Spike 铁蛋白纳米颗粒疫苗抗原进行配方开发和比较研究。
DOI：
10.1101/2023.04.03.535447
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Volkin,DavidB
中科院分区：
文献类型：
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作者：
Kumru,OzanS;Sanyal,Mrinmoy;Friedland,Natalia;Hickey,John;Joshi,Richa;Weidenbacher,Payton;Do,Jonathan;Cheng,Ya-Chen;Kim,PeterS;Joshi,SangeetaB;Volkin,DavidB
开发安全有效的第二代 COVID-19 疫苗以提高负担能力和储存稳定性要求仍然是扩大全球覆盖范围的当务之急。在本报告中，我们描述了在两种不同的细胞系中生产并用铝盐佐剂（AlHydrogel，AH）配制的自组装 SARS-CoV-2 刺突铁蛋白纳米颗粒疫苗抗原（称为 DCFHP）的配方开发和比较研究。不同水平的磷酸盐缓冲液改变了抗原-佐剂相互作用的程度和强度，并且评估了这些制剂的（1）小鼠体内性能和（2）体外稳定性特征。未加佐剂的 DCFHP 产生最小的免疫反应，而加 AH 佐剂的制剂则引起大大增强的假病毒中和滴度，与吸附到 AH 上的 DCFHP 抗原的~100%、~40% 或~10%无关。然而，这些制剂的体外稳定性特性有所不同，这是通过生物物理学研究和用于测量 AH 结合抗原的 ACE2 受体结合的竞争性 ELISA 确定的。有趣的是，4°C 保存一个月后，观察到抗原性小幅增加，同时从 AH 解吸抗原的能力下降。最后，我们对 Expi293 和 CHO 细胞中产生的 DCFHP 抗原进行了可比性评估，这显示出其 N 连接寡糖谱的预期差异。尽管由不同的 DCFHP 糖型组成，但这两种制剂在关键质量属性方面高度相似，包括分子大小、结构完整性、构象稳定性、与 ACE2 受体的结合和小鼠免疫原性特征。总而言之，这些研究支持未来在 CHO 细胞中生产的 AH 佐剂 DCFHP 候选疫苗的临床前和临床开发。
The development of safe and effective second-generation COVID-19 vaccines to improve affordability and storage stability requirements remains a high priority to expand global coverage. In this report, we describe formulation development and comparability studies with a self-assembled SARS-CoV-2 spike ferritin nanoparticle vaccine antigen (called DCFHP), when produced in two different cell lines and formulated with an aluminum-salt adjuvant (Alhydrogel, AH). Varying levels of phosphate buffer altered the extent and strength of antigen-adjuvant interactions, and these formulations were evaluated for their (1)in vivoperformance in mice and (2)in vitrostability profiles. Unadjuvanted DCFHP produced minimal immune responses while AH-adjuvanted formulations elicited greatly enhanced pseudovirus neutralization titers independent of ∼100%, ∼40% or ∼10% of the DCFHP antigen adsorbed to AH. These formulations differed, however, in theirin vitrostability properties as determined by biophysical studies and a competitive ELISA for measuring ACE2 receptor binding of AH-bound antigen. Interestingly, after one month of 4°C storage, small increases in antigenicity with concomitant decreases in the ability to desorb the antigen from the AH were observed. Finally, we performed a comparability assessment of DCFHP antigen produced in Expi293 and CHO cells, which displayed expected differences in their N-linked oligosaccharide profiles. Despite consisting of different DCFHP glycoforms, these two preparations were highly similar in their key quality attributes including molecular size, structural integrity, conformational stability, binding to ACE2 receptor and mouse immunogenicity profiles. Taken together, these studies support future preclinical and clinical development of an AH-adjuvanted DCFHP vaccine candidate produced in CHO cells.