Document Type
Thesis
Date of Award
1-31-1982
Degree Name
Master of Science in Chemical Engineering - (M.S.)
Department
Chemical Engineering and Chemistry
First Advisor
Ching-Rong Huang
Second Advisor
Gordon Lewandowski
Third Advisor
John E. McCormick
Abstract
Purification of alkaline phosphatase was experimentally investigated by semi-continous parametric pumping, and by polyacrylamide gel electrophoresis.
Alkaline phosphatase is an enzyme which is ext2acted from the human placenta and contains impurities in the form of undesired proteins. The iso-electric points of the undesired proteins are close to that of the enzyme, so a driving force other than the pH is required for their separation.
Parametric pumping involves the reciprocating flow of the enzyme mixture to be separated through a bed of ion exchanger, and synchronously changing the pH and ionic strength at the same time. The variation of pH and ionic strength displaces the interphase equilibrium, and in combination with the reciprocating flow, causes preferential movements of the sorbable components oz the mixture towards one end of the bed, leading to a build up of the separation from cycle to cycle.
Electrophoresis separation is based on the preferential movement of charged particles under the influence of an electrical field. The enzyme moves slower in an electrical field than the undesired proteins, and so can be separated.
Optimal conditions were investigated for both processes, and-it was determined that electrophoresis has a higher purification factor, while parametric pumping has a larger processing rate and a more concentrated product.
Recommended Citation
Rollan, Victor Perales, "Separation of enzyme by semi-continuous [sic] parametric pumping and by polyacrylamide gel electrophoresis" (1982). Theses. 3662.
https://digitalcommons.njit.edu/theses/3662
