Optimization of Purification Protocol for Taq DNA Polymerase used in Polymerase Chain Reaction
Description
Protein purification plays a significant role in the production of therapeutic proteins,
otherwise known as protein drugs. Although 380 protein drugs are currently marketed and used to
treat a wide range of illnesses, the protocol for protein purification varies slightly amongst
pharmaceutical companies. Thus, this study was undertaken to optimize some parameters involved
in protein purification. Taq DNA polymerase was inserted into and grown in an E. coli. cell culture,
isolated using ion gradient chromatography and a fraction collector, and assayed using polymerase
chain reaction. The concentration of isopropyl-β-D-1-thiogalactopyranoside (IPTG) used to induce
the bacterial culture and the column flow rate in chromatography were tested and adjusted to
optimize the protein purification protocol and determine their effect on both the quantity of the
purified protein collected and the enzymatic activity of the purified protein. It was discovered that
an IPTG concentration of 0.1675 mM and a column flow rate of 2 mL/min resulted in the highest
protein expression. It was concluded these values optimized the protein purification protocol both
economically and in terms of production quantity.
Subject
Protein purification
Polymerase chain reaction
Column Chromatography
Taq DNA Polymerase
IPTG
Polymerase chain reaction
Column Chromatography
Taq DNA Polymerase
IPTG