Center for Human Systems Immunology, Duke University, Durham, North Carolina, 27701, USA.
Department of Surgery, Duke University, Durham, North Carolina, 27710, USA.
Gates Open Res. 2024 Jun 28;7:107. doi: 10.12688/gatesopenres.14743.1. eCollection 2023.
Label-free techniques including Surface Plasmon Resonance (SPR) and Biolayer Interferometry (BLI) are biophysical tools widely used to collect binding kinetics data of bimolecular interactions. To efficiently analyze SPR and BLI binding kinetics data, we have built a new high throughput analysis tool named the . It can be used as a package in the Mathematica scripting environment and ultilize the non-linear curve-fitting module of Mathematica for its core function. This tool can fit the binding time course data and estimate association and dissociation rate constants ( and respectively) for determining apparent dissociation constant ( ) values. The high throughput fitting process is automatic, requires minimal knowledge on Mathematica scripting and can be applied to data from multiple label-free platforms. We demonstrate that the is optimal to analyze antibody-antigen binding data acquired on Biacore T200 (SPR), Carterra LSA (SPR imaging) and ForteBio Octet Red384 (BLI) platforms. The , and values derived using very closely matched the results from the commercial analysis software provided specifically for these instruments. Additionally, the tool generates user-directed customizable results output that can be readily used in downstream Data Quality Control associated with Good Clinical Laboratory Practice operations. With the versatility in source of data input source and options of analysis result output, the high throughput analysis tool offers investigators a powerful alternative in biomolecular interaction characterization.
无标记技术包括表面等离子体共振（SPR）和生物层干涉（BLI），是广泛用于收集双分子相互作用结合动力学数据的生物物理工具。为了有效地分析 SPR 和 BLI 结合动力学数据，我们构建了一种新的高通量分析工具，名为 。它可以作为 Mathematica 脚本环境中的一个包使用，并利用 Mathematica 的非线性曲线拟合模块作为其核心功能。该工具可以拟合结合时间过程数据，并估计缔合和解离速率常数（ 和 ），以确定表观解离常数（ ）值。高通量拟合过程是自动的，对 Mathematica 脚本的知识要求最低，并且可以应用于来自多个无标记平台的数据。我们证明 是分析在 Biacore T200（SPR）、Carterra LSA（SPR 成像）和 ForteBio Octet Red384（BLI）平台上获得的抗体-抗原结合数据的最佳选择。使用 得出的 、 和 值非常接近专门为这些仪器提供的商业分析软件的结果。此外， 工具生成用户导向的可定制结果输出，可直接用于与良好临床实验室操作相关的数据质量控制的下游操作。 高通量分析工具具有数据输入源的多功能性和分析结果输出选项，为研究人员在生物分子相互作用表征方面提供了一种强大的替代方案。
