Separation and Characterization of Deamidated Isoforms in Insulin Analogue and its Underlying Mechanism

---

Original Article

Author Details : Krishnappa Mane, Koduru Srivatsa, Ashutosh Naik, Phanichand Kodali

Volume : 5, Issue : 6, Year : 2017

Article Page : 258-264

Suggest article by email

Get Permission

---

Abstract

Deamidation of proteins has remained an elusive phenomenon largely down to the gap that exists in its understanding. One of the primary reasons, is that the experimentation involved is time consuming. However, it remains one of the most important phenomenon in proteomics both in-vitro as well as in-vivo. Deamidation results in the  quick conversion of an asparagine residue to a mixture of isoaspartate and aspartate. In glutamine residues deamidation occurs at a much lower rate. Deamidation of asparagine residues is one of the most common post-translational modifications occurring in therapeutic proteins produced using recombinant DNA technology and is the major cause for degradation of bio- pharmaceuticals. Herein, we present the separation and characterization of deamidated Isoforms in a recombinantly produced Insulin analogue by ion exchange chromatography and enzymatic digestion. The deamidation site was confirmed by using state of the art technique: LC-ESI-MS-MS. Additionally secondary structural differences were observed between the unmodified and deamidated variant of the protein.

Keywords: Deamidated isoforms, Insulin analogue.

How to cite : Mane K, Srivatsa K, Naik A, Kodali P, Separation and Characterization of Deamidated Isoforms in Insulin Analogue and its Underlying Mechanism. J Pharm Biol Sci 2017;5(6):258-264

This is an Open Access (OA) journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms.