[Heparin affinity high-performance liquid chromatography as an alternative to bioassay of CS23 mutein of recombinant human basic fibroblast growth factor].
We investigated whether a chemical assay by high-performance liquid chromatography (HPLC) as an alternative to the complicated and time-consuming bioassay for CS23 mutein of recombinant human basic fibroblast growth factor (rhbFGF-CS23) using the fetal bovine heart endothelial cell line ATCC CRL 1395. Physically, chemically or enzymatically denatured rhbFGF-CS23 was subjected to heparin affinity (HA)-HPLC and the bioassay. Good agreement was observed between the results obtained by these two methods. Moreover, HA-HPLC gave much more reproducible results (RSD = 1.9%, n = 6) than the bioassay (RSD = 7.4%, n = 18). HA-HPLC is therefore a simple, accurate and reproducible alternative to the bioassay for quality control and stability studies for rhbFGF-CS23 preparations. HA-HPLC is also considered to be applicable to assays for FGFs which have heparin affinity and biological activity similar to those of the CS23 mutein.
Hiroshima
T
T
Drug Analysis & Pharmacokinetics Research Laboratories, Takeda Chemical Industries, Ltd., Osaka, Japan.
Higashi
A
A
Imamoto
T
T
Kusumoto
S
S
Itakura
K
K
Nishijima
K
K
Nishi
K
K
D000818:Animals; D001681:Biological Assay; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016222:Fibroblast Growth Factor 2 / Q000032:analysis*; D006493:Heparin; D006801:Humans; D011994:Recombinant Proteins / Q000032:analysis
D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D006493:Heparin