Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 3584 CG, Utrecht, The Netherlands.
Department of Medical Microbiology, Amsterdam Institute for Infection and Immunity, Amsterdam UMC, University of Amsterdam, 1105AZ, Amsterdam, The Netherlands.
Chembiochem. 2022 Aug 17;23(16):e202200236. doi: 10.1002/cbic.202200236. Epub 2022 Jun 20.
Small molecule adjuvants are attractive for enhancing broad protection and durability of immune responses elicited by subunit vaccines. Covalent attachment of an adjuvant to an immunogen is particularly attractive because it simultaneously delivers both entities to antigen presenting cells resulting in more efficient immune activation. There is, however, a lack of methods to conjugate small molecule immune potentiators to viral glycoprotein immunogens without compromising epitope integrity. We describe herein a one-step enzymatic conjugation approach for the covalent attachment of small molecule adjuvants to N-linked glycans of viral glycoproteins. It involves the attachment of an immune potentiator to CMP-Neu5AcN by Cu(I)-catalyzed azide-alkyne 1,3-cycloaddition followed by sialyltransferase-mediated transfer to N-glycans of a viral glycoprotein. The method was employed to modify a native-like HIV envelope trimer with a Toll-like receptor 7/8 agonist. The modification did not compromise Env-trimer recognition by several broadly neutralization antibodies. Electron microscopy confirmed structural integrity of the modified immunogen.
小分子佐剂在增强亚单位疫苗引起的广泛保护和耐久性方面具有吸引力。将佐剂共价连接到免疫原上特别有吸引力，因为它同时将两者递送到抗原呈递细胞，从而更有效地激活免疫。然而，缺乏将小分子免疫增强剂与病毒糖蛋白免疫原缀合而不损害表位完整性的方法。本文描述了一种一步酶促偶联方法，用于将小分子佐剂共价连接到病毒糖蛋白的 N 连接糖上。它涉及通过 Cu（I）催化的叠氮化物-炔烃 1,3-环加成将免疫增强剂附着到 CMP-Neu5AcN 上，然后通过唾液酸转移酶介导转移到病毒糖蛋白的 N-糖上。该方法用于用 Toll 样受体 7/8 激动剂修饰天然样 HIV 包膜三聚体。修饰不会损害几种广泛中和抗体对 Env-三聚体的识别。电子显微镜证实了修饰后的免疫原的结构完整性。
