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Rapid Quantification of Monoclonal Antibody Titer in Cell Culture Harvests by Antibody-Induced Z-ELP-E2 Nanoparticle Cross-Linking
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-11-24 00:00:00 , DOI: 10.1021/acs.analchem.8b04083 Andrew R. Swartz 1 , Wilfred Chen 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-11-24 00:00:00 , DOI: 10.1021/acs.analchem.8b04083 Andrew R. Swartz 1 , Wilfred Chen 1
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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.
中文翻译：
通过抗体诱导的Z-ELP-E2纳米粒子交联快速定量细胞培养收获物中的单克隆抗体滴度
现有的用于定量单克隆抗体（mAb）细胞培养效价的测定方法通常需要昂贵的仪器或试剂，并且可能会受到低通量或繁琐协议的限制。在这里，我们开发了一种基于mAb诱导的与SpyTag / SpyCatcher共轭功能化的Z-domain-ELP-E2纳米笼的交联反应的快速且经济高效的替代分析。在将具有固定纳米颗粒浓度的mAb样品混合10分钟后，我们发现通过600 nm吸光度测得的浊度显示出高的信号与背景之比，并且与mAb浓度成正比。发现一个简单的对数回归拟合（R 2= 0.99）100至1000μg/ mL之间的mAb浓度的浊度数据。使用两个工业mAb细胞培养物收获物验证了优化的测定程序，并且使用Octet生物层干涉仪和Protein A传感器进行的桥接研究证实了准确且可重复的结果。该测定程序可以轻松适应高通量形式，以快速进行mAb滴度筛选。
更新日期：2018-11-24
中文翻译：
通过抗体诱导的Z-ELP-E2纳米粒子交联快速定量细胞培养收获物中的单克隆抗体滴度
现有的用于定量单克隆抗体（mAb）细胞培养效价的测定方法通常需要昂贵的仪器或试剂，并且可能会受到低通量或繁琐协议的限制。在这里，我们开发了一种基于mAb诱导的与SpyTag / SpyCatcher共轭功能化的Z-domain-ELP-E2纳米笼的交联反应的快速且经济高效的替代分析。在将具有固定纳米颗粒浓度的mAb样品混合10分钟后，我们发现通过600 nm吸光度测得的浊度显示出高的信号与背景之比，并且与mAb浓度成正比。发现一个简单的对数回归拟合（R 2= 0.99）100至1000μg/ mL之间的mAb浓度的浊度数据。使用两个工业mAb细胞培养物收获物验证了优化的测定程序，并且使用Octet生物层干涉仪和Protein A传感器进行的桥接研究证实了准确且可重复的结果。该测定程序可以轻松适应高通量形式，以快速进行mAb滴度筛选。
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