Purification Experiment Set-Up

Last Friday (October 25th), JP introduced me to another project that our lab is working on. Previously, we had two different ways to produce free peptide. First, we could grow peptides off cellulose and dissolve the cellulose in DMSO to leave the free pepide in solution. Second, we could grow peptides off of a polymer bead that has  functional groups for the peptide to attach to. These linkers could then be cut using a strong acid to produce free peptide. Now, we are working on producing peptide that grows directly off of the polymer bead, so it cannot be cleaved. Using these beads in a column, we could purify biologic. When biologic is entered into the top of the column, it would be captured by the column through the peptides. We could then elute off the biologic.

To prep for this experiment, I inserted seven specific amino acid sequences into the peptide machine program, copying each 13 times. After inserting the sequences, I used the program to see what volume of each amino acid prepped amino acids for the peptide machine. I then rounded those values up to either 5, 10, or 15 mL of solution. After determining the amount of each solution we would need, I calculated the gram amount of amino acid and mL amount of NMT to be added to make each solution. I then labeled each amino acid tube and added the specified grams of amino acid to each.

We will use the amino acid solutions that I made today to produce the seven specified peptides that I entered into the program. I look forward to returning to RPI next Friday to see what progress has been made!