The results did not support my hypothesis for the biological activity of Cone. There are some sources of error that could explain the results obtained. It’s possible there was a problem with either the Cone buffer or the sheep red blood cells to allow for all wells to turn pink and appear agglutinated.
Another explanation of the irregular results was there might have been cross contamination from not changing tips when transferring to different Cone concentrations, or if bubbles were introduced while diluting the Cone, making he results difficult to interpret. For wells A, B D, and E as Cone became more diluted or decreased in concentration, it became more difficult for it to effectively crosslink and agglutinate the red blood cells.
Well D, the positive control that contained the purchased Cone resulted in agglutination of the first couple wells, then no agglutination as the Cone concentration decreased, similar to Row A. Wells B and E that had the Glaciates additive obtained the same titer of the control Cone because Cone does not bind Calaboose. Calaboose doesn’t interfere with Cone from binding to the sugar residues on the red blood cells. Manses on the other hand, is an inhibitor to Conn’s binding sites. The Manses in solution competed with the Cone and did not allow to bind to the sugar residues on the red blood cells as seen in rows C and F.
Row G, the negative control, should have resulted in non-agglutination, similar to the rows containing the Manses additive. The results observed showed agglutination formed in this row. Lastly, Row H should have shown Nan-agglutination through out because the well contained only Cone buffer, not Cone protein. In conclusion, the results did not clearly explain the biological activity of Cone with the humiliation’s assay. The experiment contained too many anomalies to get a clear determination of Conn’s functionality post purification.
The results did show that a change in the concentration of Cone would alter the strength of the reaction. Also, Conn’s ability to bind to sugar residues can be affected if Cone has to compete or is inhibited to bind to a cells membrane.