The Use of RP-HPLC for measuring activation and cleavage of rFVIIa during purification

I Mollerup; S Bayne; E Halkjaer; T Jørgensen; G Mandrup; E M Nicolaisen; L Thim

doi:10.1002/bit.260480512

The Use of RP-HPLC for measuring activation and cleavage of rFVIIa during purification

Biotechnol Bioeng. 1995 Dec 5;48(5):501-5. doi: 10.1002/bit.260480512.

Authors

I Mollerup¹, S Bayne, E Halkjaer, T Jørgensen, G Mandrup, E M Nicolaisen, L Thim

Affiliation

¹ Biopharmaceuticals Division, Novo Nordisk A/S, Gentofte, Denmark.

PMID: 18623514
DOI: 10.1002/bit.260480512

Abstract

A reverse phase HPLC (RP-HPLC) method for analysis of recombinant factor VIIa (rFVIIa) has been developed. The method discriminates between different forms of recombinant FVII (rFVII). To obtain separation of these closely related molecules the method has been optimized with respect to gradient profile and temperature. The method has been used for optimization of purification processes and for kinetic studies. EVidence for autolytic cleavage was obtained. (c) 1995 John Wiley & Sons, Inc.