Al-Jaber, Hend Sultan A.S.
(2013)
Recombinant Erythropoietin
Extraction By Immunoaffinity Column
And The Detection Of Isoforms Using
Capillary Electrophoresis.
Masters thesis, Universiti Sains Malaysia.
Abstract
Erythropoietin (EPO) is an endogenous glycoprotein hormone. It is
synthesized in and secreted mostly by the kidneys. In the market, there are a number of
pharmaceuticals of recombinant EPO (rEPO) for clinical use. With the advent of novel
technologies in the pharmaceutical and therapeutic areas, recombinant EPO have been
introduced into the market. Although recombinant EPO has been broadly used to treat
various forms of anaemia, it has also become a very well known drug due to its misuse
in endurance sports. Sport authorities have prohibited the use of rEPO since 1989. This
work has used a number of techniques to purify natural EPO and the recombinant form
from the urine. Two Immunoaffinity column techniques (IAC) were used to purify
EPO i.e. HiTrap Affinity Columns and MAIIA EPO Purification Kit. Both columns
were successfully able to trap EPO molecules. EPO recovery from the HiTrap columns
was negligible < 1%, whereas the recovery of MAIIA affinity column was satisfactory.
The work has also developed a novel characterization tool using Capillary
Electrophoresis (CE) that can be used in pharmaceutical Quality Control (QC) and has
been investigated for its ability to detect drug misuse in sport. Conventional CE tools
generally use a label that attaches to the EPO for identification.
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