Date of Award

5-2018

Document Type

Honors Thesis

Degree Name

Bachelor of Science

Department

Biological Science

First Advisor

Trent Gemmill

Second Advisor

Susan Sharfstein

Abstract

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.

Included in

Biology Commons

Share

COinS