Formulation development of a stable influenza recombinant neuraminidase vaccine candidate.

IF 4.1 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Human Vaccines & Immunotherapeutics Pub Date : 2024-12-31 Epub Date: 2024-03-18 DOI:10.1080/21645515.2024.2304393
Bing Li, Irina V Ustyugova, Lisa Szymkowicz, Shaolong Zhu, Marin Ming, Karen Y Y Fung, Guadalupe Cortés, D Andrew James, Michael Hrynyk, Nausheen Rahman, Roger H Brookes, Salvador F Ausar
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Abstract

Current influenza vaccines could be augmented by including recombinant neuraminidase (rNA) protein antigen to broaden protective immunity and improve efficacy. Toward this goal, we investigated formulation conditions to optimize rNA physicochemical stability. When rNA in sodium phosphate saline buffer (NaPBS) was frozen and thawed (F/T), the tetrameric structure transitioned from a "closed" to an "open" conformation, negatively impacting functional activity. Hydrogen deuterium exchange experiments identified differences in anchorage binding sites at the base of the open tetramer, offering a structural mechanistic explanation for the change in conformation and decreased functional activity. Change to the open configuration was triggered by the combined stresses of acidic pH and F/T. The desired closed conformation was preserved in a potassium phosphate buffer (KP), minimizing pH drop upon freezing and including 10% sucrose to control F/T stress. Stability was further evaluated in thermal stress studies where changes in conformation were readily detected by ELISA and size exclusion chromatography (SEC). Both tests were suitable indicators of stability and antigenicity and considered potential critical quality attributes (pCQAs). To understand longer-term stability, the pCQA profiles from thermally stressed rNA at 6 months were modeled to predict stability of at least 24-months at 5°C storage. In summary, a desired rNA closed tetramer was maintained by formulation selection and monitoring of pCQAs to produce a stable rNA vaccine candidate. The study highlights the importance of understanding and controlling vaccine protein structural and functional integrity.

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稳定型流感重组神经氨酸酶候选疫苗的配方开发。
可以通过加入重组神经氨酸酶(rNA)蛋白抗原来增强目前的流感疫苗,从而扩大保护性免疫并提高疗效。为此,我们研究了优化 rNA 理化稳定性的配制条件。当磷酸钠生理盐水缓冲液(NaPBS)中的 rNA 被冷冻和解冻(F/T)时,四聚体结构会从 "封闭 "构象转变为 "开放 "构象,从而对功能活性产生负面影响。氢氘交换实验确定了开放四聚体底部锚定结合位点的差异,为构象改变和功能活性降低提供了结构机理解释。酸性 pH 值和 F/T 值的双重压力引发了开放构型的变化。所需的封闭构象在磷酸二氢钾缓冲液(KP)中得以保留,最大限度地降低了冷冻时的 pH 值下降,并加入了 10%的蔗糖以控制 F/T 应力。在热应力研究中进一步评估了稳定性,通过酶联免疫吸附试验(ELISA)和尺寸排阻色谱法(SEC)很容易检测到构象的变化。这两项测试都是稳定性和抗原性的合适指标,被认为是潜在的关键质量属性(pCQA)。为了解更长期的稳定性,对 6 个月热应激 rNA 的 pCQA 曲线进行了建模,以预测在 5°C 储存条件下至少 24 个月的稳定性。总之,通过配方选择和 pCQA 监测,可保持理想的 rNA 封闭四聚体,从而生产出稳定的 rNA 候选疫苗。这项研究强调了了解和控制疫苗蛋白质结构和功能完整性的重要性。
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来源期刊
Human Vaccines & Immunotherapeutics
Human Vaccines & Immunotherapeutics BIOTECHNOLOGY & APPLIED MICROBIOLOGY-IMMUNOLOGY
CiteScore
7.90
自引率
8.30%
发文量
489
审稿时长
3-6 weeks
期刊介绍: (formerly Human Vaccines; issn 1554-8619) Vaccine research and development is extending its reach beyond the prevention of bacterial or viral diseases. There are experimental vaccines for immunotherapeutic purposes and for applications outside of infectious diseases, in diverse fields such as cancer, autoimmunity, allergy, Alzheimer’s and addiction. Many of these vaccines and immunotherapeutics should become available in the next two decades, with consequent benefit for human health. Continued advancement in this field will benefit from a forum that can (A) help to promote interest by keeping investigators updated, and (B) enable an exchange of ideas regarding the latest progress in the many topics pertaining to vaccines and immunotherapeutics. Human Vaccines & Immunotherapeutics provides such a forum. It is published monthly in a format that is accessible to a wide international audience in the academic, industrial and public sectors.
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