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

[31] Determination of Picomolar Concentrations of Paraoxon in Human Urine by Fluorescence-Based Enzymatic Assay
Cetrangolo, Giovanni Paolo
Gori, Carla
Rusko, Janis
Terreri, Sara
Manco, Giuseppe
Cimmino, Amelia
Febbraio, Ferdinando
SENSORS, 2019, 19 (22)
[32] Superior performance of liposomes over enzymatic amplification in a high-throughput assay for myoglobin in human serum
Edwards, Katie A.
Meyers, Katherine J.
Leonard, Barbara
Baeumner, Antje J.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2013, 405 (12) : 4017 - 4026
[33] A Fluorescence-Based High-Throughput Screen to Identify Small Compound Inhibitors of the Genotype 3a Hepatitis C Virus RNA Polymerase
Eltahla, Auda A.
Lackovic, Kurt
Marquis, Christopher
Eden, John-Sebastian
White, Peter A.
JOURNAL OF BIOMOLECULAR SCREENING, 2013, 18 (09) : 1027 - 1034
[34] A general fluorescence-based coupled assay for S-adenosylmethionine-dependent methyltransferases
Wang, CH
Leffler, S
Thompson, DH
Hrycyna, CA
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2005, 331 (01) : 351 - 356
[35] A homogeneous, high-throughput fluorescence resonance energy transfer-based DNA polymerase assay
Shapiro, A
Rivin, O
Gao, N
Hajec, L
ANALYTICAL BIOCHEMISTRY, 2005, 347 (02) : 254 - 261
[36] Fluorescence-based assay probing regulator of G protein signaling partner proteins
Huang, Po-Shiun
Yeh, Hsin-Sung
Yi, Hsiu-Ping
Lin, Chain-Jia
Yang, Chii-Shen
ANALYTICAL BIOCHEMISTRY, 2012, 423 (01) : 133 - 140
[37] Identification of a lead small-molecule inhibitor of anthrax lethal toxin by using fluorescence-based high-throughput screening
Wei, Dong
Bu, Zhaoyun
Yu, Ailian
Li, Feng
BMB REPORTS, 2011, 44 (12) : 811 - 815
[38] High-throughput microfluidic devices for NO assay based on electrophoretic separation
Miyado, T
Tanaka, Y
Nagai, H
Takeda, S
Saito, K
Fukushi, K
Yoshida, Y
Wakida, S
Niki, E
2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology, 2005, : 66 - 68
[39] Development of fluorescence-based high throughput screening assays: choice of appropriate instrumentation
Burns, D
Alder, L
Fan, YH
McKeegan, E
Warrior, U
Beutel, B
SYSTEMS AND TECHNOLOGIES FOR CLINICAL DIAGNOSTICS AND DRUG DISCOVERY, PROCEEDINGS OF, 1998, 3259 : 188 - 196
[40] A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction
Fan, Xiaoqin
Wei, Jinmei
Xiong, Haiting
Liu, Xiaohui
Benichou, Serge
Gao, Xuejuan
Liu, Langxia
INTERNATIONAL JOURNAL OF ONCOLOGY, 2015, 47 (04) : 1485 - 1493

← 1 2 3 4 5 →