A Fluorescence-Based High-Throughput Coupled Enzymatic Assay for Quantitation of Isoaspartate in Proteins and Peptides

被引：6
作者：
Puri, Aastha [1 ]
Quan, Yong [1 ]
Narang, Ajit S. [1 ]
Adams, Monica [1 ]
Gandhi, Rajesh [1 ]
Nashine, Vishal C. [1 ]
机构：
[1] Bristol Myers Squibb Co, Drug Prod Sci & Technol, 1 Squibb Dr, New Brunswick, NJ 08901 USA
来源：
AAPS PHARMSCITECH | 2017年 / 18卷 / 03期
关键词：
deamidation; fluorescence; high-throughput; isoaspartate; protein; stability; ADENOSYLMETHIONINE-DEPENDENT METHYLTRANSFERASES; MASS-SPECTROMETRY ANALYSIS; COLORIMETRIC ASSAY; DEAMIDATION; ACID;
D O I：
10.1208/s12249-016-0570-7
中图分类号：
R9 [药学];
学科分类号：
1007 ;
摘要：
Formation of isoaspartate (IsoAsp) from spontaneous asparagine (Asn) deamidation or aspartate (Asp) isomerization is one of the most common non-enzymatic pathways of chemical degradation of protein and peptide pharmaceuticals. Rapid quantitation of IsoAsp formation can enable rank-ordering of potential drug candidates, mutants, and formulations as well as support shelf life prediction and stability requirements. A coupled enzymatic fluorescence-based IsoAsp assay (CEFIA) was developed as a high-throughput method for quantitation of IsoAsp in peptides and proteins. In this note, application of this method to two therapeutic candidate proteins with distinct structural scaffolds is described. In addition, the results obtained with this method are compared to those from conventional assays.
引用
收藏
页码：803 / 808
页数：6
相关论文
共 50 条
  • [41] A High-Throughput Bioluminescent Assay to Monitor the Deamidation of Asparagine and Isomerization of Aspartate Residues in Therapeutic Proteins and Antibodies
    Hsiao, Kevin
    Alves, Juliano
    Patel, Rushikesh
    Adams, Monica
    Nashine, Vishal
    Goueli, Said
    JOURNAL OF PHARMACEUTICAL SCIENCES, 2017, 106 (06) : 1528 - 1537
  • [42] Evaluation of the effects of 18 non-synonymous single-nucleotide polymorphisms of CYP450 2C19 on in vitro drug inhibition potential by a fluorescence-based high-throughput assay
    Wang, Huijuan
    Kim, Raymond A.
    Sun, Dongxu
    Gao, Yiwen
    Wang, Hao
    Zhu, Juanli
    Chen, Chao
    XENOBIOTICA, 2011, 41 (09) : 826 - 835
  • [43] Quantification of Membrane Protein Self-Association with a High-Throughput Compatible Fluorescence Assay
    Li, Junbei
    Qiu, Xiaoyan J.
    BIOCHEMISTRY, 2017, 56 (14) : 1951 - 1954
  • [44] Assay establishment and validation of a high-throughput organoid-based drug screening platform
    Xiaomeng Li
    Guoxiang Fu
    Long Zhang
    Ruoyu Guan
    Peiyuan Tang
    Jialing Zhang
    Xinxin Rao
    Shengzhi Chen
    Xiaoya Xu
    Yi Zhou
    Yun Deng
    Tao Lv
    Xingfeng He
    Shaobo Mo
    Peiyuan Mu
    Jianjun Gao
    Guoqiang Hua
    Stem Cell Research & Therapy, 13
  • [45] Assay establishment and validation of a high-throughput organoid-based drug screening platform
    Li, Xiaomeng
    Fu, Guoxiang
    Zhang, Long
    Guan, Ruoyu
    Tang, Peiyuan
    Zhang, Jialing
    Rao, Xinxin
    Chen, Shengzhi
    Xu, Xiaoya
    Zhou, Yi
    Deng, Yun
    Lv, Tao
    He, Xingfeng
    Mo, Shaobo
    Mu, Peiyuan
    Gao, Jianjun
    Hua, Guoqiang
    STEM CELL RESEARCH & THERAPY, 2022, 13 (01)
  • [46] High-throughput assay of tyrosine phenol-lyase activity using a cascade of enzymatic reactions
    Zhu, Hang-Qin
    Hu, Wen-Ye
    Tang, Xiao-Ling
    Zheng, Ren-Chao
    Zheng, Yu-Guo
    ANALYTICAL BIOCHEMISTRY, 2022, 640
  • [47] Superior performance of liposomes over enzymatic amplification in a high-throughput assay for myoglobin in human serum
    Katie A. Edwards
    Katherine J. Meyers
    Barbara Leonard
    Antje J. Baeumner
    Analytical and Bioanalytical Chemistry, 2013, 405 : 4017 - 4026
  • [48] Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay
    Marina A. Kalinina
    Dmitry A. Skvortsov
    Maria P. Rubtsova
    Ekaterina S. Komarova
    Olga A. Dontsova
    Molecular Imaging and Biology, 2018, 20 : 368 - 377
  • [49] Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay
    Kalinina, Marina A.
    Skvortsov, Dmitry A.
    Rubtsova, Maria P.
    Komarova, Ekaterina S.
    Dontsova, Olga A.
    MOLECULAR IMAGING AND BIOLOGY, 2018, 20 (03) : 368 - 377
  • [50] Fluorescence-Based High Throughput Screening Technologies for Natural Chloride Ion Channel Blockers
    Xu, Rong
    Xiao, Yuan
    Liu, Yan
    Wang, Bo
    Li, Xing
    Huo, Cong
    Jia, Xin
    Hou, Liming
    Wang, Xiaoming
    CHEMICAL RESEARCH IN TOXICOLOGY, 2018, 31 (12) : 1332 - 1338
12345