What is Fmoc deprotection?

Fmoc (9-fluorenylmethoxycarbony-) group is the most commonly N-terminal protecting group used in Solid Phase Peptide Synthesis (SPPS) (Scheme 1, Table 1). Furthermore, the Fmoc deprotection step is one of the most crucial stages in peptide synthesis (besides amino acids coupling).

What is Fmoc used for?

The fluorenylmethoxycarbonyl protecting group (Fmoc) is a base-labile protecting group used in organic synthesis.

Why is Fmoc used in peptide synthesis?

In Fmoc solid-phase peptide synthesis, the peptide chain is assembled stepwise, one amino acid at a time, while attached to an insoluble resin support. Fmoc synthesis is mild, flexible and versatile and consequently offers more synthetic options than the alternative Boc chemistry.

What is Fmoc SPPS?

Today, Fmoc SPPS is the method of choice for peptide synthesis. Very-high-quality Fmoc building blocks are available at low cost because of the economies of scale arising from current multiton production of therapeutic peptides by Fmoc SPPS.

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.

How do I remove trityl protecting group?

Trityl (triphenylmethyl, Tr) – Removed by acid and hydrogenolysis. Silyl ether (most popular ones include trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), tri-iso-propylsilyloxymethyl (TOM), and triisopropylsilyl (TIPS) ethers) – Removed by acid or fluoride ion.

Which pair is best utilized for the protection of amines?

Mate! The most popular choice of protecting group for amine nitrogen is the carbamate functional group.

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.

What is the longest peptide synthesized using SPPS?

The longest peptide we have made to date is 169 amino acids. We promise to provide the HPLC purified peptide with correct molecular weight analyzed by Electrospray ionization (ESI) Mass Spectrometry.

What is Edman degradation?

Edman degradation is the process of purifying protein by sequentially removing one residue at a time from the amino end of a peptide. To solve the problem of damaging the protein by hydrolyzing conditions, Pehr Edman created a new way of labeling and cleaving the peptide.

What is a coupling reagent?

Benzotriazole-based peptide coupling reagents and additives, as activators or additives (Figure 1), are used in peptide bond formation reactions both in solution and solid-phase synthesis. There are many different types of peptide coupling reagents (e.g., carbodiimides, aminium/uranium salts, and phosphonium salts).

How is Aspartimide formation prevented?

One of the simplest strategies for limiting aspartimide formation is to change the Fmoc-removal conditions. Simply adding 0.1 M hydroxybenzotriazole (HOBt) to the piperidine solution has been shown to reduce aspartimide formation significantly.

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 reagent for Fmoc deprotection?

Table 2: Properties of reagents 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).

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.

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. 1.