Solid Phase Peptide Synthesis Mechanism

The processing of peptide is referred to as peptide synthesis. Over the course of the year, numerous processes and methods for manufacturing large quantities of peptides were discovered and patented in order to satisfy the protein’s demand in various medical fields. Organic chemistry has made a significant contribution to the mechanism of peptide synthesis. visit
Peptide synthesis is reliable and error-free. However, there are a few things that can severely jeopardise the protocols’ reproducibility. The consistency of DMF is perhaps the most troubling of all the variables. To achieve a higher yield, it is critical to use ‘quality’ DMF during solid phase peptide synthesis. This entails either removing it from the solvent system or starting over with a new container. There are only a few solid phase peptide synthesis pathways that fall under this group.
The first step in solid-phase peptide synthesis is to decide what functional group your C-terminus should be:
Using 2-chlorotrityl resin if you want the C-terminus to be a carboxylic acid.
Using Rink amide resin if you want your C-terminus to be an amide.
Using 2-chlorotrityl resin when producing macrocyclic peptides.
After you’ve decided on a resin, you’ll need to load the first amino acid onto it.
1- Measuring an acceptable volume of resin is the first step. For a 0.1 mmol scale synthesis, 300 mg is typically used. In a Poly-Prep chromatography panel, unload the resin (BioRad).
2- Allow resin to swell in CH2Cl2 at room temperature for at least 30 minutes (longer is fine).
3- Weigh out the first amino acid and dissolve it in 8 mL CH2Cl2 with 0.3 mL 2,4,6-collidine. Our first amino acid in a macrocyclic peptide is almost always Boc-Orn(Fmoc)-OH. Using approximately 100 mg Boc-Orn(Fmoc)-OH.
4- Extract all CH2Cl2 from the column containing the swelled resin and substitute it with the Amino acid/DCM/Collidine solution using a nitrogen gas flow.
5- Rock for a minimum of 8 hours (no longer than 24 hours).
6. Finish capping the 2-chlorotrityl Resin.
Resin 2-Cholotrityl Capping
This phase is necessary to covalently attach a small nucleophile (methanol) to unreacted carbocations on the 2-chlorotrityl chloride resin.