方案摘要
方案下载应用领域 | 生物产业 |
检测样本 | 其他 |
检测项目 | |
参考标准 | / |
本应用中描述了CD光谱法在高浓度抗体(未稀释)中的扩展应用,并报告了使用Spectra Manager Ver.2.5 CFR Bestsel对抗体药物进行详细二级结构评估的结果。
Higher-order structure (HOS) is an important physicochemical property of antibody drugs. The most well-known method for HOS characterization is circular dichroism ( CD ) spectroscopy . Althougl CD spectroscopy can provide robust structural information for low-concentration protein solutions ( 0.1 mg / ml ) , antibody drugs are formulated at concentrations higher than 10 mg / ml Under these conditions , the antibody molecules can exhibit physicochemical properties that differ from those under dilute conditions . Fortunately , evaluation of high concentration solutions can be done via short CD path lengths ( 10 mm ) or using FTIR with ATR . The ability to perform structural characterization of antibodies without dilution is extremely important because intermolecular interactions and changes in the HOS can lead to the formation of aggregates that cause serious Immunogenicity.
Secondary structure estimation offers a route to evaluate protein higher order structure , but historically has not worked well on proteins with high amounts of b-sheet such as antibodies.Recently , Micsonai et al developed the Bestsel algorithm that can accurately estimate the secondary structure from a CD spectrum by taking into account parallel-antiparallel orientations of b-strands and the twist of antiparallel b-sheets . Bestsel has the following features : 1 ) high estimation accuracy for a wide range of proteins , including b-strand-rich proteins such as antibodies , 2 ) provides eight types of secondary structure information , 3 ) capabiility to predict protein folding following the CATH classification , and 4 ) open web server .
While many academic researchers use the Bestsel web server , researchers in biopharma companies who need to work in a GXP environment have not been able to benefit from Bestsel . To make Bestsel accessible to biopharma , we developed Spectra Manager Ver.2.5 CFR Bestsel as an add-in software for Spectra Manager , a control and analysis platform for JASCO CD spectrometers , which is compatible with GXP.
Here , we describe an expanded application of CD spectroscopy for highly concentrated antibodies (undiluted) and report the results of a detailed secondary structure evaluation of antibody drugs using Spectra Manager Ver.2.5 CFR Bestsel.
本应用中结论:
J-1500 CD光谱仪和Spectra ManagerTM2.5 BeStSel CFR能够跟踪配制抗体药物的详细结构变化。
Spectra Manager TM2.5 BeStSel CFR提供符合GxP的环境,并提供符合ALCOA+原则的CSV、ER/ES和Dl。
HPLC 中的采样频率、响应时间和柱外效应
基于抗体的药物强制降解研究中聚集物和降解产物的SEC监测
用CD光谱法评估VHH抗体的高阶结构、稳定性和相似性
相关产品
JASCO圆二色光谱仪CD J-1500
JASCO振动圆二色光谱仪VCD FVS-6000
JASCO圆偏振荧光光谱仪CPL-300
JASCO高效色谱仪LC-4000
jascoRFT-6000傅立叶变换红外拉曼光谱仪
JASCO红外显微镜IRT-7200
MSV-5000显微紫外可见近红外分光光度计
jasco紫外可见分光光度计 V-700
JASCO FTIR-4000傅立叶变换红外光谱仪
JASCO 全自动数字旋光仪P-2000
JASCOFP-8000系列荧光光谱仪
JASCO SFC4000超临界流体色谱仪
JASCONRS5000/7000共聚焦激光拉曼光谱仪
JASCO V-7000系列紫外可见近红外分光光度计
JASCO傅立叶变换红外光谱仪FT/IR-6000
关注
拨打电话
留言咨询