Monitoring biological purification

In the discovery and development of hylan A and hylan B, of course, the most important criteria were their purity and biocompatibility. We could never have used these two molecules as therapeutics without the availability of the test that I developed in the late 1960s to monitor the purification process for the native hyaluronan, and producing NIF-NaHA. This test method was accepted by many regulatory agencies as a standard way of proving the biological compatibility of elastoviscous fluids used in ophthalmic... 

HPLC has played an important role for years in the drug discovery process in pharmaceutical laboratories. HPLC has proven a valuable asset in purifications of drug from de novo synthesis, from biological matrices, and from combinational synthesis. HPLC assay has moved from the discovery laboratory on through manufacturing, production, and metabolite monitoring. 

The biological activity was monitored throughout the isolation stages, i.e., from solvent extraction to final HPLC purification. The 10 mg of material was further purified to give 8 mg of crystalline material. On the basis of the yield of the crystalline product, the active ingredient was shown to be present at a level of 200 ppb in pollen. 

One of the critical factors in our research has been the adaptation and use of multiple assays to follow heparinase activity. Particularly important were assays (e.g., Azure A) used in monitoring the fermentation and early stages of purification. By utilizing three different approaches for assaying heparin (disappearance of heparin, appearance of reaction products, and disappearance of heparin s biological activity), the occurrence of any arti-... 

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