What is solution phase peptide synthesis?

Abstract. Solution phase synthesis was the first developed and the only method for peptide synthesis until the solid phase peptide synthesis (SPPS) introduced by Merrifield revolutionized the way peptides and their analogues are prepared nowadays.

How do you Deprotect Fmoc?

Standard Removal of Fmoc Protecting Group

1. Place the resin in a round bottom flask and add 20% (v/v) piperidine in DMF (approximately 10 mL/gm resin).
2. Shake the mixture at room temperature for 2 minutes.
3. Filter the resin.
4. Add a second portion of 20% piperidine in DMF.
5. Shake the mixture at room temperature for 5 minutes.

What is liquid phase peptide synthesis?

Liquid-phase peptide synthesis is a classical approach to peptide synthesis. It has been replaced in most labs by solid-phase synthesis (see below). However, it retains usefulness in large-scale production of peptides for industrial purposes.

How can Epimerization be prevented?

To reduce the epimerization in chiral molecules, you can try your chiral molecule with a substrate stereospecific (i.e. Rutenium complex for the hydrogenation of geranil molecules). This substrate will give you and unique product, with high abundance.

Which is a main advantage of solid phase synthesis?

General advantages of solid phase synthesis are easy purification, rapid generation of linear peptide intermediates, and precedent in the synthesis of large peptides.

Is Fmoc stable to acid?

The Fmoc group is acid stable and Boc-Lys(Fmoc)-OH is used to prepare protected peptide fragments for fragment coupling.

What is liquid phase synthesis?

Liquid phase synthesis includes the low evaporation, hydrothermal/solvothermal treatments, electrochemical/sonochemical protocols, microwave, spray-drying, flow chemistry and ionothermal methods.

What are the steps in peptide synthesis?

First an amino acid is coupled to the resin. Subsequently, the amine is deprotected, and then coupled with the free acid of the second amino acid. This cycle repeats until the desired sequence has been synthesized. SPPS cycles may also include capping steps which block the ends of unreacted amino acids from reacting.

How do I get rid of Fmoc group?

The Fmoc group is, in general, rapidly removed by primary (i.e., cyclohexylamine, ethanolamine) and some secondary (i.e., piperidine, piperazine) amines, and slowly removed by tertiary (i.e., triethylamine [Et3N], N, N-diisopropylethylamine [DIEA]) amines.

How does HOBt prevent racemization?

Adding HOBt, 6-Cl-HOBt or HOAt suppresses the racemization. Protecting the pi imidazole nitrogen in the histidine side-chain with the methoxybenzyl group greatly reduces racemization.

Why is Fmoc deprotection important in peptide synthesis?

Furthermore, the Fmoc deprotection step is one of the most crucial stages in peptide synthesis (besides amino acids coupling). Most importantly, the property which makes the Fmoc group a valuable tool in SPPS is its selective base-mediated removal while leaving the other, acid-labile side-chain protecting groups intact.

Which is the best solvent for Fmoc deprotection?

Solvents for Fmoc Deprotection The Fmoc removal reaction is usually performed in polar electron donor solvents: dimethylformamide (DMF) or N-methylpirrolidone (NMP). However, DMF and NMP do not have a high potential to disrupt the interchain aggregations (like TFA has).

Which is the best way to remove the Fmoc group?

Use of N – (2-mercaptoethyl)aminomethyl polystyrene resin as the dibenzofulvene scavenger provided a convenient means of deblocking the Fmoc group by simple filtration and evaporation to the free base. These methods are an improvement over conventional methods and should find a broad utility for deblocking Fmoc-protected amines.

How is the deprotection of Fmoc-Phe-AMC performed?

First, the deprotection of Fmoc-Phe-AMC using DBU and 10 equivalents of either thiophenol, DTT, and octanethiol was monitored for unreacted DBF by HPLC.