Development, methodological evaluation and application of a cell-based TRF assay for analysis of ADCC activity
文献类型：期刊论文
| 作者 | Zhu, Xiao1,2; Gong, Likun1,2 ; Qin, Qiuping2
|
| 刊名 | JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
 |
| 出版日期 | 2023-10-25 |
| 卷号 | 235页码:7 |
| ISSN号 | 0731-7085 |
| 关键词 | Antibody-dependent cell-mediated cytotoxicity Time-resolved fluorescence Monoclonal antibody Antibody-Drug Conjugate Eu NK-92 cells |
| DOI | 10.1016/j.jpba.2023.115655 |
| 通讯作者 | Gong, Likun(lkgong@cdser.simm.ac.cn) ; Qin, Qiuping(qpqin@cdser.simm.ac.cn) |
| 英文摘要 | Interaction of an antibody with its Fc & gamma;R plays an important role in effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC). Nowadays altered ADCC activity of an antibody can be achieved by utilizing an effective glyco-engineering strategy, which often involves changes of sugar moieties in Fc part of the antibody, thereby affecting its receptor binding with effector cells. We aimed to construct a cell-based time-resolved fluorescence (TRF) assay for the evaluation of ADCC activity triggered by the antibody drug Trastuzumab (antiHER2) and T-DM1. The assay was initiated by incubating 2,2 & PRIME;:6 & PRIME;,2 "-Terpyridine-6,6"-dicarboxylic acid (TDA)labeled target SK-BR3 cells with the testing antibodies and engineered NK-92 effector cells. After incubation, the target cells were lysed to detect TDA released into the supernatant. Together with added Eu, the TDA in the supernatant formed a stable chelate of EuTDA with high-intensity fluorescence. The ADCC activity was then determined by measuring the fluorescence of EuTDA. Consequently, the method demonstrated good accuracy, precision, linearity, and specificity over methodological assessment and compared well with the Luciferase release assay in terms of the agreement of the achieved results. Using the developed assay, we evaluated the ADCC activity of two glyco-engineered anti-HER-2 antibody-drug conjugates (ADCs) and the results showed that antibody Fc glycosylation modifications influenced antibody ADCC activity to varying degrees. In conclusion, the present assay is able to accurately assess the ADCC activity induced by Trastuzumab (anti-HER2) and T-DM1, and a similar methodology can be applied to other therapeutic antibodies during drug development to help screen for antibodies with desirable ADCC activity. |
| WOS关键词 | FC-GAMMA-RIIIA ; IN-VITRO ; ANTIBODIES ; IGG |
| WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| 出版者 | ELSEVIER |
| WOS记录号 | WOS:001067661200001 |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/306149]  |
| 专题 | 中国科学院上海药物研究所 |
| 通讯作者 | Gong, Likun; Qin, Qiuping |
| 作者单位 | 1.Nanjing Univ Chinese Med, Nanjing 210023, Jiangsu Provinc, Peoples R China 2.Chinese Acad Sci, Shanghai Inst Mat Med, Ctr Drug Safety Evaluat & Res, Dept Immunoassay & Immunochem, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhu, Xiao,Gong, Likun,Qin, Qiuping. Development, methodological evaluation and application of a cell-based TRF assay for analysis of ADCC activity[J]. JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS,2023,235:7. |
| APA | Zhu, Xiao,Gong, Likun,&Qin, Qiuping.(2023).Development, methodological evaluation and application of a cell-based TRF assay for analysis of ADCC activity.JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS,235,7. |
| MLA | Zhu, Xiao,et al."Development, methodological evaluation and application of a cell-based TRF assay for analysis of ADCC activity".JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 235(2023):7. |
入库方式： OAI收割
来源：上海药物研究所
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