Group of Molecular Virology, Centro Singular de Investigación en Química Biolóxica e Materiais Moleculares (CiQUS), Departamento de Bioquímica e Bioloxía Molecular, Universidade de Santiago de Compostela, Santiago de Compostela, 15782, Spain.
Centro Singular de Investigación en Química Biolóxica e Materiais Moleculares (CIQUS), Departamento de Química Orgánica, Universidade de Santiago de Compostela, Santiago de Compostela, 15782, Spain.
Sci Rep. 2018 Nov 2;8(1):16286. doi: 10.1038/s41598-018-34488-3.
We have previously developed a methodology to produce protein microspheres (MS) that can be loaded with proteins of interest in living cells through their C or N-terminal tagging with the so-called IC-Tag. The IC-Tagging method has many applications ranging from the production of immobilized enzymes for industrial use to the production of subunit vaccines due to its intrinsic adjuvancy. Here we show the adaptation of the IC-Tagging to work inside the endoplasmic reticulum and bacteria, allowing us to produce properly modified viral glycoproteins. Additionally, we were able to express the Islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP), whose expression remained elusive to date possibly due to its toxicity when over-expressed. IGRP is an antigen of enormous pharmaceutical interest as it is specifically targeted during the autoimmune response taking place in both the Non-Obese Diabetic (NOD) mice and type 1 diabetes (T1D) patients leading to the destruction of insulin-producing beta cells.
我们之前开发了一种方法来生产蛋白质微球 (MS),通过所谓的 IC 标签,可以将其 C 或 N 末端与感兴趣的蛋白质连接,从而将其加载到活细胞中。IC 标记法具有许多应用,从生产用于工业用途的固定化酶到生产亚单位疫苗,因其内在的佐剂作用。在这里,我们展示了 IC 标记法在真核细胞和细菌内工作的适应性,使我们能够生产出适当修饰的病毒糖蛋白。此外,我们能够表达胰岛特异性葡萄糖-6-磷酸酶催化亚基相关蛋白 (IGRP),其表达至今仍难以捉摸,可能是因为其在过表达时具有毒性。IGRP 是一种具有巨大药物价值的抗原,因为它在非肥胖型糖尿病 (NOD) 小鼠和 1 型糖尿病 (T1D) 患者中发生的自身免疫反应中被特异性靶向,导致产生胰岛素的β细胞被破坏。