Structural changes of interfacially adsorbed antibody molecules

界面吸附抗体分子的结构变化

• 批准号: BB/S018492/1
• 负责人: Jian Lu
• 金额: $ 29.04万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FS018492%2F1
• 关键词: Structural; changes; interfacially; adsorbed; antibody

Cancer treatments have undergone three revolutionary stages over the past few decades: chemotherapies, biomarkers targeted at mutated genes, and combined treatments of biomarker targeting and immune process mediation. Each stage of these treatments was facilitated by advances in our understanding of the behaviour of cancers, especially at the molecular and cell levels. Our early understanding of the high growth rates of cancerous cells led to the development of chemical/drug therapies together with radiation treatments to kill cancerous cells. However, such medical treatments must undergo rigorous clinical trials and meet regulatory requirements, e.g., FDA approval, before clinical deployment. This process plus further improvement to perfect the treatments can take more than a decade. Currently, chemotherapy is still the mainstream cancer treatment for patients, but their toxicity remains a major limiting factor to survival rates. Over the past 5-8 years, the ability to moderate immune processes has been realised, and a number of pioneering treatments based on this new line of thinking have very recently achieved clinical success. This prospect has driven a major global effort to develop new treatments based on antibody technologies, covering not only oncology but also other major diseases including cardiovascular, respiratory, autoimmunity and infectious diseases. Antibodies used for cancer or other disease treatments must be designed, manufactured, separated, purified and eventually formulated into medical products ready for clinical use. A popular means of administration is to apply via intravenous injection. This option requires the antibody drugs to be formulated as a stable protein solution in a bottle (glass or plastic) or a ready-to-inject syringe set, with a shelf-life between 1-2 years. Because these bioengineered antibodies have to be equipped with two or more biological functions, their amino acid sequences (so called primary sequences) must be altered. As a result, we do not know how stable their folded domains are and how instability from the modified domains will affect the stability of the whole antibody.All proteins are amphiphilic due to the presence of both polar and apolar amino acids on their surfaces. This amphiphilic character drives proteins to adsorb and desorb at different interfaces spontaneously. During these interfacial processes, proteins interact with the substrate surface and with themselves, and depending on the nature of the substrate surface and the close proximity between them once adsorbed, deformation of the globular structures and even local unfolding can occur, causing exposure of hydrophobic patches that may induce aggregation and precipitation, compromising the bioactivity of such antibody drugs. Newly bioengineered antibodies are often unstable, and adsorption can accelerate their instability.Using a series of bioengineered antibodies with well-controlled sequence modifications in Fab and Fc domains, this LINK project forges a new collaborative team involving MedImmune, Manchester University and Imperial College London, with the aim to develop new understanding by combining neutron reflection experiments with molecular dynamics simulation. We will examine how certain well-controlled sequence modifications in Fab and Fc domains affect their adsorbed globular structures and how instability in these domains affects the structure of the whole mAb. Another line of work will be to examine how representative substrate surfaces affect structural deformation and unfolding. These studies will lead to new results that will be of great value to the biopharmaceutical industry and to academic research. The successful delivery of this project will lead to new primary structure-stability relationships that will assist MedImmune and other protein drug developers to improve their antibody stability in their biotherapeutics. The outcome will ultimately benefit the general public.

在过去的几十年里，癌症治疗经历了三个革命性的阶段：化疗，靶向突变基因的生物标志物，以及生物标志物靶向和免疫过程介导的联合治疗。这些治疗的每个阶段都得益于我们对癌症行为的理解，特别是在分子和细胞水平上的理解。我们对癌细胞高生长率的早期理解导致了化学/药物疗法以及放射治疗的发展，以杀死癌细胞。然而，这种医学治疗必须经过严格的临床试验并满足监管要求，例如，FDA批准，在临床部署之前。这个过程加上进一步改进以完善治疗可能需要十多年。目前，化疗仍然是患者的主流癌症治疗方法，但其毒性仍然是生存率的主要限制因素。在过去的5-8年里，人们已经实现了调节免疫过程的能力，并且基于这种新思维的许多开创性治疗方法最近已经取得了临床成功。这一前景推动了一项重大的全球努力，即开发基于抗体技术的新疗法，不仅涵盖肿瘤学，还涵盖其他主要疾病，包括心血管、呼吸道、自身免疫和传染病。用于癌症或其他疾病治疗的抗体必须经过设计、制造、分离、纯化并最终配制成可供临床使用的医疗产品。一种流行的给药方式是通过静脉注射施用。这种选择需要将抗体药物配制成瓶（玻璃或塑料）或即用注射器套件中的稳定蛋白质溶液，保质期在1-2年之间。由于这些生物工程抗体必须具备两种或更多种生物学功能，因此必须改变其氨基酸序列（所谓的一级序列）。因此，我们不知道它们的折叠结构域有多稳定，以及修饰结构域的不稳定性如何影响整个抗体的稳定性。所有蛋白质都是两亲性的，这是由于它们表面上存在极性和非极性氨基酸。这种两亲性特征驱动蛋白质自发地在不同界面吸附和解吸。在这些界面过程中，蛋白质与基底表面和自身相互作用，并且取决于基底表面的性质和它们之间一旦被吸附的紧密接近度，可能发生球状结构的变形甚至局部解折叠，导致疏水性斑块的暴露，这可能诱导聚集和沉淀，损害此类抗体药物的生物活性。新的生物工程抗体通常不稳定，吸附会加速其不稳定性，利用一系列在Fab和Fc结构域进行了良好控制的序列修饰的生物工程抗体，这个LINK项目组建了一个新的合作团队，包括MedImmune，曼彻斯特大学和伦敦帝国理工学院伦敦，旨在通过结合中子反射实验和分子动力学模拟来开发新的理解。我们将研究Fab和Fc结构域中某些控制良好的序列修饰如何影响其吸附的球状结构，以及这些结构域中的不稳定性如何影响整个mAb的结构。另一项工作将是研究代表性的基底表面如何影响结构变形和展开。这些研究将导致新的结果，将是非常有价值的生物制药行业和学术研究。该项目的成功交付将导致新的一级结构-稳定性关系，这将有助于MedImmune和其他蛋白质药物开发人员提高其生物治疗中的抗体稳定性。其结果最终将使广大公众受益。

项目成果

How do Self-Assembling Antimicrobial Lipopeptides Kill Bacteria?

• DOI: 10.1021/acsami.0c17222
• 发表时间: 2020-12-16
• 期刊: ACS APPLIED MATERIALS & INTERFACES
• 影响因子: 9.5
• 作者: Gong, Haoning;Sani, Marc-Antoine;Lu, Jian Ren
• 通讯作者: Lu, Jian Ren

In-Membrane Nanostructuring of Cationic Amphiphiles Affects Their Antimicrobial Efficacy and Cytotoxicity: A Comparison Study between a De Novo Antimicrobial Lipopeptide and Traditional Biocides.

• DOI: 10.1021/acs.langmuir.2c00506
• 发表时间: 2022-05-31
• 期刊: LANGMUIR
• 影响因子: 3.9
• 作者: Fa, Ke;Liu, Huayang;Gong, Haoning;Zhang, Lin;Liao, Mingrui;Hu, Xuzhi;Ciumac, Daniela;Li, Peixun;Webster, John;Petkov, Jordan;Thomas, Robert K.;Lu, Jian Ren
• 通讯作者: Lu, Jian Ren

What happens when pesticides are solubilised in binary ionic/zwitterionic-nonionic mixed micelles?

• DOI: 10.1016/j.jcis.2020.10.083
• 发表时间: 2021-03-15
• 期刊: JOURNAL OF COLLOID AND INTERFACE SCIENCE
• 影响因子: 9.9
• 作者: Hu, Xuzhi;Gong, Haoning;Lu, Jian R.
• 通讯作者: Lu, Jian R.

Structural Disruptions of the Outer Membranes of Gram-Negative Bacteria by Rationally Designed Amphiphilic Antimicrobial Peptides

• DOI: 10.1021/acsami.1c01643
• 发表时间: 2021-04-02
• 期刊: ACS APPLIED MATERIALS & INTERFACES
• 影响因子: 9.5
• 作者: Gong, Haoning;Hu, Xuzhi;Lu, Jian Ren
• 通讯作者: Lu, Jian Ren

Acidification-Induced Structure Evolution of Lipid Nanoparticles Correlates with Their In Vitro Gene Transfections.

酸化诱导的脂质纳米颗粒的结构演变与其体外基因转染相关。

• DOI: 10.1021/acsnano.2c06213
• 发表时间: 2023-01-06
• 期刊: ACS NANO
• 影响因子: 17.1
• 作者: Li, Zongyi;Carter, Jessica;Santos, Luis;Webster, Carl;Walle, Christopher F. van der;Li, Peixun;Rogers, Sarah E.;Lu, Jian Ren
• 通讯作者: Lu, Jian Ren