Solid-phase peptide synthesis [Video file].

One of the most often asked questions is what region of a protein to synthesize to produce anti-peptide antisera. Although there is no correct answer for any protein, there are a number of computer programs derived from several publications that provide good approximations for immunogenic regions within proteins . These are available from the Rockefeller University Computer Services in the Smith Hall User Area.

Prior to finalizing your peptide sequence please consider the following points:

Synthesis of difficult peptide sequences: A comparison …

Stephen Kent on Solid-phase peptide synthesis, part of a collection of online lectures.

Synthesis of difficult peptide sequences: a comparison …

81. Bacsa B, Horvati K, Bosze S. . Solid-phase synthesis of difficult peptide sequences at elevated temperatures: a critical comparison of microwave and conventional heating technologies. 2008;73:7532-42

Overview of Solid Phase Synthesis of “Difficult Peptide” Sequences

80. Palasek SA, Cox ZJ, Collins JM. Limiting racemization and aspartimide formation in microwave-enhanced Fmoc solid phase peptide synthesis. 2007;13:143-8

T1 - Solid-phase synthesis of difficult peptide sequences at elevated temperatures

Stephen Kent on Solid-phase peptide synthesis, ..

Each peptide sequence is unique and some peptides sequences prove problematic during synthesis. If problems do occur with your order we will inform you as soon as possible. Sometimes peptides have to be resynthesised to provide the required yield, meaning orders will take longer to complete. In these instances part of your order will be shipped as soon as it is ready, and the remaining amount will be supplied as soon as possible.

'Difficult' peptide sequences in SPPS;

(Chemical Equation Presented) The Fmoc/t-Bu solid-phase synthesis of three difficult peptide sequences (a 9-mer, 15-mer, and 24-mer) was performed using N,N′-diisopropylcarbodiimide/1-hydroxybenzotriazole as coupling reagent on polystyrene, Tentagel, and ChemMatrix resins. In order to obtain an insight into the specific role of the elevated temperature and/or the electromagnetic field for peptide syntheses carried out using microwave irradiation, peptide couplings and Fmoc-deprotection steps were studied under microwave and conventionally heated conditions at the same temperature. While room temperature couplings/deprotections generally produced the difficult peptides in rather poor quality, excellent peptide purities were obtained using microwave heating at a temperature of 86°C for both the coupling and deprotection steps in only 10 and 2.5 min reaction time, respectively. While for most amino acids no significant racemization was observed, the high coupling temperatures led to considerable levels of racemization for the sensitive amino acids His and Cys. It was demonstrated for all three peptide sequences that when performing the coupling/deprotection steps at the same reaction temperature using conventional heating, nearly identical results in terms of both peptide purity and racemization levels were obtained. It therefore appears that the main effect of microwave irradiation applied to solid-phase peptide synthesis is a purely thermal effect not related to the electromagnetic field.

(1990) Prediction of difficult sequences in solid phase peptide synthesis.

Custom Peptide Synthesis Services ..

88(3), Johnson EC, Kent SB, Exploratory synthesis of peptide-alpha-thioester segments spanning the polypeptide sequence of the delta-opioid receptor, a G protein-coupled receptor, 340-9, 2007 John Wiley and Sons, Inc." class="disabled">

14. Moss JA. Guide for resin and linker selection in solid-phase peptide synthesis.  2005 Chapter 18: Unit 18.7

Peptide synthesis services for research ..

56. Dolphin GT. A designed well-folded monomeric four-helix bundle protein prepared by Fmoc solid-phase peptide synthesis and native chemical ligation. 2006;12:1436-47

In liquid phase peptide synthesis, the “difficult sequence” is closely related to the solubility of protected peptides.

Solid-phase Synthesis of Difficult Peptides and ..

Since the purity of the peptide obtained from the synthesis is sequence dependent, purification to >95% will be performed upon request. Major impurities can range from 10% to 50% by weight. They consist of small water-soluble molecules, salts and protecting groups from the cleavage reaction, deletion peptides created due to incomplete coupling during synthesis, and modified peptides created during the cleavage. These species can be removed using reversed phase HPLC. We recommend that, under most circumstances, all peptides be purified prior to employing them in research studies.