Solid Phase Peptide Synthesis Mechanism

The processing of peptide is referred to as peptide synthesis. Over the course of the year, various processes and methods for producing large numbers 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. Click next to learn more.

Peptide synthesis is reliable and error-free. However, there are a few things that can seriously jeopardise the protocols’ reproducibility. The consistency of DMF is perhaps the most troubling of all the factors. 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 category.

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. Use approximately 100 mg Boc-Orn(Fmoc)-OH.

4- Remove all CH2Cl2 from the column containing the swelled resin and replace 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.