Antibody engineering to identify optimal expression systems and glycoforms of IgE class antibodies for cancer therapy

癌症治疗的设计策略主要集中在九种人类抗体中的一种，IgG1。我们设计了一种替代方法，通过工程抗癌抗体携带IgE Fc类区域。已知IgE抗体支持对寄生虫的免疫防御，并与过敏有关。这些活性可能作为一种新的癌症免疫治疗策略被利用来对抗癌症，例如，我们的一流IgE抗体识别肿瘤相关抗原，正在盖伊医院进行I期临床试验。IgE是一种高度糖基化的抗体，但其糖基部分对IgE抗癌的效力和免疫细胞激活机制的确切影响尚不清楚。在这项研究中，我们的目标是探索最佳工程和糖工程策略，以产生抗肿瘤IgE类抗体，以确定免疫肿瘤学的最佳抗体。我们将生成在不同哺乳动物表达系统中表达相同特异性的IgEs，并将表征其与功能相关的碳水化合物谱。此外，我们将通过施用聚糖生物合成抑制剂产生不同的IgE糖型（Wagner）。抗体变异产生和药理学评估将在第一年进行。第2-3年将重点研究抗体在患者血液和血清中的生物学功能、效力和体外安全性研究。此外，糖谱分析（Ludger Ltd）将进行并与抗体功能谱相关。该研究旨在帮助鉴定具有明确功能谱的IgE抗体和糖变异体，并具有相当大的潜力，为肿瘤临床试验提供优化的抗体免疫疗法。项目目标：我们将运用我们已经建立的表达克隆方法，利用糖基转移酶抑制剂在不同的哺乳动物表达系统和不同的糖型中生产抗肿瘤IgE抗体克隆。我们将询问抗体变体的功能和糖基化谱。一组具有明确属性的特异性抗体将被选择用于下游翻译和临床测试。

Design strategies for cancer therapy have largely focused on one of nine human antibody classes, IgG1. We have designed an alternative approach by engineering anti-cancer antibodies bearing IgE Fc class regions. IgE antibodies are known to support immune defences against parasites and to be associated with allergies. These activities may be harnessed against cancer as a novel cancer immunotherapy strategy, exemplified by our first-in-class IgE antibody recognising a tumour-associated antigen undergoing a Phase I clinical trial at Guy's Hospital. IgEs are highly glycosylated antibodies, yet the precise effects of IgE glycan moieties on the potency and immune cell-activating mechanisms of IgE against cancer remain unclear. In this study, we aim to explore optimal engineering and glycoengineering strategies for the generation of anti-tumour IgE class antibodies in order to identify optimised antibodies for immuno-oncology. We will generate IgEs of the same specificity expressed in different mammalian expression systems and we will characterise their carbohydrate profiles in relation to function. Furthermore, we will generate different IgE glycoforms by administering inhibitors of glycan biosynthesis (Wagner). Antibody variant generation and pharmacological evaluations will be conducted in Year 1. Years 2-3 will focus on elucidating antibody biological functions, potency and ex vivo safety studies in patient blood and sera. Furthermore, glycoprofiling analyses (Ludger Ltd) will be conducted and correlated with antibody functional profiles. The study is designed to aid the identification of IgE antibodies and glycovariants with defined functional profiles and harbours considerable potentialto deliver optimised antibody immunotherapies for clinical testing in oncology.Project Aims: We will apply our established expression cloning methodologies to produce anti-tumour IgE antibody clones in different mammalian expression systems and as different glycoforms using glycosyltransferase inhibitors. We will interrogate the functional and glycosylation profiles of antibody variants. A panel of characterised antibodies with defined attributes will be selected for downstream translation and clinical testing.