Chemical and Biomolecular Engineering, Department of


Date of this Version



A THESIS Presented to the Faculty of The Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Master of Science, Major: Chemical Engineering, Under the Supervision of Professor William H. Velander. Lincoln, Nebraska: August, 2012

Copyright (c) 2012 Ayman Ismail


A solvent detergent treated fibrinogen was purified from human plasma by cryoprecipitation (cryo) followed by chemical precipitation using ethanol (EtOH) or ammonium sulfate (AS) as precipitating agents. Amounts of fibronectin (FN), factor XIII A-subunit (FXIIIA), factor XIII b-subunit (FXIIIB), and alpha2-antiplasmin (α2-AP) in the isolated fibrinogen were quantified. Thromboelastography (TEG) analysis was used to evaluate the clot strength of the isolated fibrinogen and to determine the ability of the ethanol and ammonium sulfate precipitations to eliminate the solvent detergent. Sodium dodecylsulfate-polyacrylamide gel analysis indicated that fibrinogen produced by each of these precipitation methods had similar purity. Quantitative western blot analysis revealed that fibrinogen produced by ammonium sulfate precipitation contained increased amounts of FN, FXIIIB, and α2-AP. TEG analysis showed that ammonium sulfate precipitated fibrinogen yielded a fibrin clot with the highest maximal strength. In addition, a single ethanol precipitation was sufficient to remove the solvent detergent while a single ammonium sulfate precipitation was not effective in removing the solvent detergent mixture Tri (n-butyl) phosphate (TNBP) and Triton X-100 as judged by the ability of the material to form a clot.

Adviser: William H. Velander