Week 6 - Parker

Research Project: Prostate Cancer Antibody

This week I have been spending the majority of my time on my research project. I am working on quantifying the internalization of the J591 antibody in LnCap cells, a prostate cancer cell line. The antibody I am testing is actually a single chain version of the monoclonal antibody. I have been having some issues with labeling the antibody using the amines, this is due to the size of the single chain. There are a lot fewer amines for the fluorophore to conjugate to. However, the antibody has a his-tag located at the n-terminal end of the protein that could be exploited to label the antibody.

We are now attempting to label the single chain using a new conjugation method that binds the fluorophore to the his-tag instead of the amines. We hope that this will allow for enough conjugation to reduce the noise we are seeing with the current method. In addition, we hope that the single chain will have results comparable to what we are seeing with the monoclonal antibody staining. Thus far we have been unsuccessful in comparing the two due to the poor conjugation we are seeing with the single chain.

The significance of this work is in the unique targeting and production of a targeting agent. The single chain will be easier to make than the monoclonal antibody and could possibly allow for mass production of a targeting agent for prostate cancer therapies.

S. Parker