Rapid Quantification of Monoclonal Antibody Titer in Cell Culture Harvests by Antibody-Induced Z-ELP-E2 Nanoparticle Cross-Linking
Rapid Quantification of Monoclonal Antibody Titer in Cell Culture Harvests by Antibody-Induced Z-ELP-E2 Nanoparticle Cross-Linking
复制标题
通过抗体诱导的 Z-ELP-E2 纳米颗粒交联快速定量细胞培养收获物中的单克隆抗体滴度
DOI：
10.1021/acs.analchem.8b04083
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发表时间：
2018
影响因子：
7.4
通讯作者：
Chen, Wilfred
中科院分区：
文献类型：
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作者：
Swartz, Andrew R.;Chen, Wilfred
现有的单克隆抗体（mAb）细胞培养滴度的定量分析通常需要昂贵的仪器或试剂，并且可能受到低通量或繁琐方案的限制。在这里，我们开发了一种基于单克隆抗体诱导的与SpyTag/SpyCatcher偶联功能化的z -domain elp - e2纳米笼交联的快速且具有成本效益的替代方法。将mAb样品与固定的纳米颗粒浓度混合10分钟后，我们发现浊度在600 nm处通过吸光度测量，表现出高信号与背景比，并与mAb浓度成正比。单抗浓度在100 ~ 1000 μg/mL范围内，浊度数据经简单对数回归拟合（R2= 0.99）。优化的分析程序通过两次工业单克隆抗体细胞培养收获进行了验证，并使用蛋白a传感器的八tet生物层干涉法进行桥接研究，证实了准确和可重复的结果。分析程序可以很容易地适应快速单抗滴度筛选的高通量格式。
Existing assays for the quantification of monoclonal antibody (mAb) cell culture titer often require expensive instruments or reagents and may be limited by the low-throughput or tedious protocols. Here, we developed a quick and cost-effective alternative assay based on mAb-induced cross-linking with Z-domain-ELP-E2 nanocages functionalized by SpyTag/SpyCatcher conjugation. After mixing mAb samples with a fixed nanoparticle concentration for 10 min, we found that the turbidity, measured by absorbance at 600 nm, exhibited a high-signal-to-background ratio and was proportional to the mAb concentration. A simple logarithmic regression was found to fit (R2= 0.99) the turbidity data for mAb concentrations between 100 and 1000 μg/mL. The optimized assay procedure was validated using two industrial mAb cell culture harvests, and a bridging study using Octet biolayer interferometry with Protein A sensors confirmed accurate and reproducible results. The assay procedure can be easily adapted to a high-throughput format for rapid mAb titer screening.