Development of a Rapid Small-Scale Purification Method for the Quantitation of Heparin-Like Glycosaminoglycans from Cell Culture Media
Date of Award
Bachelor of Science
Heparin is an anticoagulant medicinally used to inhibit blood clotting. It is commonly administered to patients requiring surgery or kidney dialysis. Presently, it is produced from animal tissues, but a recent contamination crisis pointed to the need for a safer source of the drug. Our project seeks to develop a rapid, inexpensive, high-throughput assay to quantitate the production of heparin and other glycosaminoglycans (GAGs) from cultured mammalian cells. In order to quantitate heparin from cell culture media, a purification method is needed to separate GAGs from interfering constituents in the media. We developed a purification protocol that absorbs Pluronic and most proteins to Sep-Pak C-18 cartridges, followed by ultrafiltration through 5 kDa cutoff Vivaspin centrifugal concentrators. The purified GAGs are then quantitated by a microcarbazole assay.
Infantado, Marina Danielle, "Development of a Rapid Small-Scale Purification Method for the Quantitation of Heparin-Like Glycosaminoglycans from Cell Culture Media" (2018). Biological Sciences. 49.