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

页码：803 / 808

页数：6

共 50 条

[41] A fluorescence-based high-throughput screening method for cytokinin translocation mutants
Zhang, Mengyuan
Ding, Bingli
Zhao, Jiangzhe
Zhang, Penghong
Li, Yujia
Yang, Guodong
Zhang, Kewei
PLANT METHODS, 2020, 16 (01)
[42] Substrate arrays for fluorescence-based enzyme fingerprinting and high-throughput screening
Reymond, Jean-Louis
FLUORESCENCE METHODS AND APPLICATIONS: SPECTROSCOPY, IMAGING, AND PROBES, 2008, 1130 : 12 - 20
[43] Recent Advances in Design of Fluorescence-Based Assays for High-Throughput Screening
Fang, Xiaoni
Zheng, Yongzan
Duan, Yaokai
Liu, Yang
Zhong, Wenwan
ANALYTICAL CHEMISTRY, 2019, 91 (01) : 482 - 504
[44] A fluorescence-based high-throughput screening method for cytokinin translocation mutants
Mengyuan Zhang
Bingli Ding
Jiangzhe Zhao
Penghong Zhang
Yujia Li
Guodong Yang
Kewei Zhang
Plant Methods, 16
[45] A robust fluorescence-based assay for human erythrocyte Ca++ efflux suitable for high-throughput inhibitor screens
Sims, Jeremiah N.
Yun, EJun
Chu, Jonathan
Siddiqui, Mansoor A.
Desai, Sanjay A.
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS, 2023, 52 (1-2): : 101 - 110
[46] A robust fluorescence-based assay for human erythrocyte Ca++ efflux suitable for high-throughput inhibitor screens
Jeremiah N. Sims
EJun Yun
Jonathan Chu
Mansoor A. Siddiqui
Sanjay A. Desai
European Biophysics Journal, 2023, 52 : 101 - 110
[47] Novel High-Throughput Fluorescence-Based Assay for the Identification of Nematocidal Compounds That Target the Blood-Feeding Pathway
Marchand, Anthony
Van Bree, Joyce W. M.
Taki, Aya C.
Moyat, Mati
Turcatti, Gerardo
Chambon, Marc
Smith, Adam Alexander Thil
Doolan, Rory
Gasser, Robin B.
Harris, Nicola Laraine
Bouchery, Tiffany
PHARMACEUTICALS, 2022, 15 (06)
[48] A Fluorescence-Based High-Throughput Assay for the Discovery of Exchange Protein Directly Activated by Cyclic AMP (EPAC) Antagonists
Tsalkova, Tamara
Mei, Fang C.
Cheng, Xiaodong
PLOS ONE, 2012, 7 (01):
[49] 5-Hydroxymethylcytosine as a clinical biomarker: Fluorescence-based assay for high-throughput epigenetic quantification in human tissues
Margalit, Sapir
Avraham, Sigal
Shahal, Tamar
Michaeli, Yael
Gilat, Noa
Magod, Prerna
Caspi, Michal
Loewenstein, Shelly
Lahat, Guy
Friedmann-Morvinski, Dinorah
Kariv, Revital
Rosin-Arbesfeld, Rina
Zirkin, Shahar
Ebenstein, Yuval
INTERNATIONAL JOURNAL OF CANCER, 2020, 146 (01) : 115 - 122
[50] Development of a Fluorescence-Based, High-Throughput SARS-CoV-2 3CLpro Reporter Assay
Froggatt, Heather M.
Heaton, Brook E.
Heaton, Nicholas S.
JOURNAL OF VIROLOGY, 2020, 94 (22)

← 1 2 3 4 5 →