HEPARIN AFFINITY HIGH-PERFORMANCE LIQUID- CHROMATOGRAPHY AS AN ALTERNATIVE TO BIOASSAY OF CS23 MUTEIN OF RECOMBINANT HUMAN BASIC FIBROBLASTGROWTH-FACTOR

We investigated whether a chemical assay by high-performance liquid ch romatography (HPLC) as an alternative to the complicated and time-cons uming bioassay for CS23 mutein of recombinant human basic fibroblast g rowth factor (rhbFGF-CS23) using the fetal bovine heart endothelial ce ll line ATCC CRL 1395. Physically, chemically or enzymatically denatur ed rhbFGF-CS23 was subjected to heparin affinity (HA)-HPLC and the bio assay. Good agreement was observed between the results obtained by the se 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 bioa ssay for quality control and stability studies for rhbFGF-CS23 prepara tions. HA-HPLC is also considered to be applicable to assays for FGFs which have heparin affinity and biological activity similar to those o f the CS23 mutein.