This week began very slowly, with Monday and Tuesday being spent entirely working on the computer. During this time I reviewed last week’s experiment procedures and results and made sure that we had protocols and visuals that would allow for reliable results and reproducibility. I also updated the procedure for the the first bead conjugation that was performed on Wednesday.
On Wednesday I was able to perform the first bead conjugation of the summer. This process is likely the main source of failures for the project. So to combat this, I performed it at OraSure under the instruction and supervision of one of their experienced scientists. This allowed me to further learn the ins and outs of the procedure, as well as valuable techniques that may not be in the protocol, but will lead to better more consistent results. This experiment in particular was to determine the optimal pH for our conjugation to occur at using two differently pHed buffers. The experiment was successful. We ran test strips comparing the performance of the two differently conjugated beads and saw that there was a clear better option. This will allow us to move forward, tweaking other aspects of our protocol until we are able to reliably produce excellent results. These results felt like a huge step forward, we confirmed a successful conjugation and have the steps in place to further optimize that process.
During this procedure another possible experiment arose in conversation. OraSure encouraged us to find every aspect of the protocol that could be challenged and major places where our protocol differed from the manufacturer’s. One of the largest differences in the two protocols was a set of additional washes that may either help with bead performance or cause particle aggregation, rendering the beads less effective. Following this discussion, I confirmed that this was something my team would be interested in evaluating. Since it was I wrote the protocol that would allow us to evaluate this through performance on test strips coupled with particle analysis at OraSure.
Another goal of this week was to make it possible to recreate every aspect of experiments performed at OraSure in the HST, our home lab. We are able to do this by utilizing some equipment that has been present, but never used by our team before. This includes and incubator and nanodrop spectrophotometer. The latter requires us to learn how to properly use it. We did this by making various dilutions of BSA (a cheap, common protein) in DI Water and we used the nanodrop to prove that the dilutions were the concentrations that we expected. We were able to do this with under 2.5% error in four different concentrations of the solution. This led to us writing a protocol for other team members to use in the future because we plan to incorporate the nanodrop into many of our future experiments. Using the nanodrop will allow us to have certainty that we have the concentration of antibodies that we think we have.
