Background::
Protein A affinity chromatography is often employed as the most crucial
purification step for monoclonal antibodies to achieve high yield with purity and throughput requirements.
Introduction::
Protein A, also known as Staphylococcal protein A (SPA) is found in the cell wall of
the bacteria staphylococcus aureus. It is one of the first discovered immunoglobulin binding
molecules and has been extensively studied since the past few decades. The efficiency of Protein A
affinity chromatography to purify a recombinant monoclonal antibody in a cell culture sample has
been evaluated, which removes 99.0% of feed stream impurities.
Materials and Method::
We have systematically evaluated the purification performance by using a
battery of analytical methods SDS-PAGE (non-reduced and reduced sample), Cation Exchange
Chromatography (CEX), Size-exclusion chromatography (SEC), and Reversed phased-Reduced
Chromatography for a CHO-derived monoclonal antibody.
Results and Discussion::
The analytical test was conducted to determine the impurity parameter,
Host Cell Contaminating Proteins (HCP). It was evaluated to be 0.015ng/ml after the purification
step; while initially, it was found to be 24.431ng/ml.
Conclusion::
The tests showed a distinct decrease in the level of different impurities after the chromatography
step. It can be concluded that Protein A chromatography is an efficient step in the purification
of monoclonal antibodies.
Attempts to obtain anti(D2 dopamine receptor) antibodies via the anti-idiotypic route
