Application Note
HTRF profiling services on SpectraMax iD5 Multi-Mode Microplate Reader
- Obtain consistent compound profiling results with the HTRF optimized SpectraMax iD5 Multi-Mode Microplate Reader
- Analyze data quickly with preconfigured protocols in SoftMax Pro Software
- Access a wide range of acclaimed HTRF assays and services with Cisbio's US Custom Solutions Laboratory team
Natalia Castro | Assay Development Scientist | Cisbio
Arturo Gonzalez Moya | Custom Solutions Manager | Cisbio
Cathy Olsen, PhD | Sr. Applications Scientist | Molecular Devices
Introduction
Cisbio’s Custom Solutions Laboratory team has worked in collaboration with Molecular Devices to generate relevant data promoting the expert profiling service activity completed by Cisbio US utilizing current Molecular Devices readers.
The SpectraMax® iD5 Multi-Mode Microplate Reader is HTRF-certified, with an enhanced TRF detection module and filters optimized for HTRF detection. Preconfigured HTRF protocols in SoftMax® Pro Software 7.0.3 and higher include optimized reader settings, as well as data analysis, for faster results.
Experimental details
A profiling service was completed using three commercial drugs all on Cisbio’s Fcγ receptor binding assays and FcRn. The SpectraMax iD5 reader was used for measurement of the results. For each drug/antibody used, Cisbio established:
- Dose-response curves in triplicate with eight (8) concentrations
- Control 1: Labeled cells expressing the receptor of interest only (min)
- Control 2: Labeled cells expressing the receptor of interest and IgG-red only (max)
Compound dose response: 8-point dose response curve of serially diluted compound performed in triplicate.
The following graphs contain the dose responses from two (2) users on two (2) different days. Data includes one single sample in all Fcγ receptors and FcRn assay (SpectraMax iD5 reader), with no outliers, bottom constrained and concentration [ng/mL].
The following graphs show Log IC50 and CV% values for all samples. Samples were run over multiple days with multiple users with readings from Molecular Devices SpectraMax iD5 Reader.
The following tables represent the Log IC50 [ng/mL] and CV% raw values from seven (7) receptors, two (2) users, and readings from the SpectraMax iD5 reader.
Conclusion
Due to the collaboration with Molecular Devices and the SpectraMax iD5 Multi-Mode Microplate Reader, Cisbio can now offer custom HTRF profiling services as a reliable, fast and reproducible approach to drug discovery using technology suitable for antibody affinity and the potential of biosimilarity studies using relevant drugs available on the market.
Natalia Castro | Assay Development Scientist | Cisbio
Arturo Gonzalez Moya | Custom Solutions Manager | Cisbio
Cathy Olsen, PhD | Sr. Applications Scientist | Molecular Devices
简介
Cisbio 定制解决方案实验室团队与 Molecular Devices 合作，使用 Molecular Devices 读 板机生成相关数据，促进 Cisbio US 完成专 家分析服务活动。SpectraMax®iD5 多功能 读板机通过 HTRF 认证，配置增强型 TRF 检测模块和针对 HTRF 检测优化的滤光片。 使用 SoftMax®Pro 7.0.3 及更高版本软件中 预先配置的 HTRF 程序模板，包括优化的 读数设置和数据分析，便于获得更快的分 析结果。
试验细节
使用三种商用药物完成一项分析服务，这 些药物均用于 Cisbio 的 Fcγ 受体结合分析 和 FcRn。在 SpectraMax iD5 读板机上测 量结果。
Cisbio 建立的对于使用的每种药物/抗体
- 剂量 - 反应曲线三个平行孔共 8 个浓度梯 度 (8)
- 对照 1：仅表达目的受体的标记细胞 ( 最 小 )
- 对照 2：仅表达目的受体和 IgG-红色荧光 的标记细胞 ( 最大 )
化合物剂量反应：化合物连续稀释 8 个浓 度点的剂量响应曲线，每个浓度下三个平 行孔。
下图展示了两 (2) 名用户在两 (2) 天内完成的 剂量反应。数据来自一个样品在所有 Fcγ 受 体和 FcRn 的测定 ( SpectraMax iD5 读板 机 )，没有异常值，浓度梯度底部约束曲线模 型 [ng / mL]
下图显示了所有样品的 Log IC50 和 CV ％ 值，样品的读数由不同的用户在 Molecular Devices 的 SpectraMax iD5 读板机上运行不同天检测获得
The following graphs show Log IC50 and CV% values for all samples. Samples were run over multiple days with multiple users with readings from Molecular Devices SpectraMax iD5 Reader.
下表显示来自七 (7) 个受体，两 (2) 个用户的 Log IC50 [ng / mL] 和 CV ％ 原始值，以及来自于 SpectraMax iD5 读板机的读数
结论
由于与 Molecular Devices 和 SpectraMax iD5 多功能读板机的合作，Cisbio 现在可提供定制的 HTRF 分析服务，作为一种可靠，快速和可重 现药物研发的方法，这个技术适用于使用抗体亲和力检测技术的药物研发，以及对市场上现有的相关药物进行生物相似性研究的潜力。