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解决FMC63-CD19亲和关系的团

Jacqueline Seigner^1,2, Charlotte U Zajc^1,3, Sarah Dötsch⁴

¹Department of Chemistry, Institute of Biochemistry, University of Natural Resources and Life Sciences, Vienna, Austria.

Scientific reports

|December 28, 2023

概括

对CD19的FMC63单链可变片段 (scFv) 亲和力被确定为2-6nM,使用优化的表面等离子体共振. 这项研究澄清了之前的CAR T细胞亲和度测量中的差异.

科学领域:

免疫学免疫学免疫学
生物化学生物化学
生物技术是生物技术.

背景情况:

针对CD19的化学抗原受体 (CAR) T细胞疗法得到了广泛的批准.
FMC63单链可变片段 (scFv) 是许多 CD19 定向的 CAR T 细胞产品中的一个关键组件.
之前的报道显示,FMC63-scFv与CD19的结合亲和力报告的变异性很大 (>100倍).

研究的目的:

为了明确确定FMC63-scFv与CD19的结合亲和力.
识别和解释之前的亲和度测量的错误来源.
为改进CAR T细胞设计和CAR亲和力-功能关系解释提供可靠的亲和力值.

主要方法:

表面等离子共振 (SPR) 用于亲和度测量.
严格单体和正确折叠的可溶性CD19被用作目标分析剂.
测试了各种实验条件,以识别和最小化导致亲和力过度或低估的文物.

主要成果:

优化的SPR实验为FMC63-scFv与CD19产生了2-6nM的结合亲和力.
低于最佳的实验设计被确定为报告的亲和关系中显著变化的原因.
阐明了以前发表的广泛亲和关系的潜在解释.

结论:

在优化条件下,FMC63-scFv与CD19的结合亲和度为2-6nM.
这项研究提供了一个验证的亲和值,对CAR T细胞研究至关重要.
这些发现将有助于解释CAR亲和力-功能关系,并设计下一代CAR T细胞.

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