A Fluorescence-Based High-Throughput Coupled Enzymatic Assay for Quantitation of Isoaspartate in Proteins and Peptides
| Autor: | Vishal Nashine, Monica L. Adams, Ajit S. Narang, Rajesh B. Gandhi, Yong Quan, Aastha Puri |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Pharmacology toxicology Mutant Pharmaceutical Science Peptide Aquatic Science 01 natural sciences Fluorescence Isoaspartate 03 medical and health sciences Isomerism Drug Discovery Asparagine Deamidation Ecology Evolution Behavior and Systematics Enzyme Assays chemistry.chemical_classification Aspartic Acid Isoaspartic Acid Chromatography Ecology 010401 analytical chemistry Proteins General Medicine 0104 chemical sciences 030104 developmental biology Enzyme chemistry Biochemistry Peptides Agronomy and Crop Science |
| Zdroj: | AAPS PharmSciTech. 18:803-808 |
| ISSN: | 1530-9932 |
| DOI: | 10.1208/s12249-016-0570-7 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink