Employ Florida Banner Center Protein Purification and Characterization Workforce Training, FSCJ

Employ Florida Banner Center Protein Purification and Characterization Workforce Training

The Banner Protein Purification workshop was held at Florida State College at Jacksonville’s Advanced Technology Center on October 29th and 30th 2012.  This was our second two day workforce training this month. It was an excellent follow up to the Cell Culture Methods course offered previously. The focus during the Protein Purification workshop was to learn and process purification of monoclonal antibodies. After purification, we measured the absorbance of our samples with a UV spectrophotometer. We proceeded to run our samples on a vertical PAGE gel and then stained for visualization.

The first day of the workshop everything went smoothly. Each group made up their PBS buffer solutions will ease. We then centrifuged ascites to remove large particles. The process of Mircofiltering the Ascites was an activity that everyone was interested in performing! I think it was the small sense of danger with working with the needle. We had to aspirate the sample into a syringe and then use the micron filters for optimal purification. We filtered our antibodies by doing numerous centrifuge rounds using the Protein A Affinity column and PBS buffers to bind our antibodies and then the Glycine elution buffer to release our samples. Our samples were then ready for refrigeration and to continue the next day.

The second day started off with some trouble shooting exercises. A student accidently chose recalibration setting instead of running his sample on the UV spectrometer. It was a great lesson on trouble shooting.  Errors are always the best learning experiences! Once the machine was reset, we set up and began running our samples in the UV spectrometer to get our standard curve.  We prepared our sample to run in the PAGE electrophoresis and learned how the chemicals in the buffer helped to denature the proteins to obtain a great separation for tracking.  The lab ended with the dying of our gels to read our results.

Lecture by Kevin R. Pegg, PhD combined with the lab, was a great way to learn how to use S.O.P’s, laboratory equipment, techniques, the protein purification process, as well as a understanding how these methods are used in real world applications.  Students asked questions and were genuinely interested in learning all the details about this process. All the attendees I spoke with thought that the workshop was great and that it would be a great benefit toward their specific goals for employment within the field and say they would love to see more of these workshops.

 

-Meagan

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