The administration of blinatumomab was accompanied by several adverse effects, including activation of regulatory T-cells and cytokine storm. The objective of this study was to produce and evaluate a novel αCD8/CD19 BiTE (αCD8/CD19) with the potency to directly target CD8⁺ T-cells. In-silico studies were utilized for determining proper folding, receptor binding, and structural stability of αCD8/CD19 protein. Western blotting and indirect surface staining were used to evaluate the size accuracy and binding potency of the purified protein. Functionality was assessed for granzyme B production, cytotoxicity, and proliferation. The αCD8/CD19 recombinant protein was produced in the CHO-K1 cell line with a final concentration of 1.94 mg/l. The αCD8/CD19 bound to CD8⁺ and CD19⁺ cell lines and induced significant granzyme B production, cytotoxic activity and proliferation potential in the presence of IL-2 and tumor target cells. The maximum CD8⁺ T-cell biological activity was observed on the 10th day with 10:1 effector-to-target ratio.

blinatumomab的施用伴随着多种不良反应，包括调节性T细胞的激活和细胞因子风暴。本研究的目的是生产和评估一种新型 αCD8/CD19 BiTE (αCD8/CD19)，其具有直接靶向 CD8 ⁺ T细胞的能力。利用计算机研究确定 αCD8/CD19 蛋白的正确折叠、受体结合和结构稳定性。使用蛋白质印迹和间接表面染色来评估纯化蛋白的大小准确性和结合效力。评估了颗粒酶 B 的产生、细胞毒性和增殖的功能。 αCD8/CD19 重组蛋白在 CHO-K1 细胞系中产生，终浓度为 1.94 mg/l。 αCD8/CD19 与 CD8 ⁺和 CD19 ⁺细胞系结合，并在 IL-2 和肿瘤靶细胞存在的情况下诱导显着的颗粒酶 B 产生、细胞毒活性和增殖潜力。第 10 天观察到最大 CD8 ⁺ T细胞生物活性，效应子与靶标比率为 10:1。