Synthesis manual peptide

The sequence of solid-phase assembly and cyclization reactions used to prepare the bis(Ampa)-bridged peptide 31 is summarized in Scheme 19. All solid-phase procedures were performed in a standard, custom-built, 200-mL capacity, manual peptide synthesis reaction vessel. 165 ... 

We have found that although the use of an automatic synthesizer can save time and can be relatively unproblematic for simple sequences, the synthesis of peptides manually allows for more flexibility and control over the assembly process. This is particularly important for the synthesis of difficult sequences as it permits quick and easy intervention at any point. 

The apparatus routinely used in this laboratory for the manual synthesis of peptides consists of a flask attached to a glass manifold that can support up to four sintered funnels, thereby permitting the simultaneous synthesis of up to four peptides. The side arm of the flask is attached to a vacuum pump to allow for solvents to be aspirated from each funnel. The manifold also contains valves... 

M Ruvo, P Scardino, G Cassani, G Fassina. Facile manual synthesis of peptide libraries. Peptide Protein Lett 1(3) 187-192, 1994. 

MBHA resin, 3.0 g (>0.2 mmol/g), is added to a filtered assembly suitable for manual peptide synthesis. DCM is added and the resin is allowed to swell for 1 h. DGM should completely cover the resin. DCM is removed by vacuum filtration and the (DCM/DIEA) solution is added to neutralize the resin. After 1 min the base solution is removed by filtration and this treatment is repeated. The resin is washed with DCM and air dried by vacuum filtration for 5 min. 

The protocols have been written as they would be carried out using a manual peptide synthesis vessel. Whilst it is appreciated that most scientists preparing peptides will be using automated peptide synthesizers, it is not possible, given the wide variation in operating procedures, to describe how such methods may be applied to individual instruments. Particular emphasis has been given here to those operations which are typically carried out off-instrument, such as first residue attachment and peptide-resin cleavage. 

Figure 1. Manual peptide synthesis bubbler vessel.

Pre-swell the peptidyl-resin (1 mmol) with DCM in a manual peptide synthesis vessel, and then wash with DCM (3 times). 

Place the peptidyl resin in a sintered glass funnel or manual peptide synthesis vessel. 

In the general procedure manual peptide synthesis is described. Kaiser test see Note 5) should be performed after each amino acid coupling ensuring completeness of the reaction. In our lab, we use a Syro II robot (MultiSynTech) for automated SPPS. If you are equipped with a robot, use your established standard protocols for peptide preparation, because the sequence is not known to be difficult. 

Equipment Single mode microwave synthesizer configured for manual peptide synthesis equipped with a fiber optic temperature probe (Discover SPS microwave-assisted peptide synthesizer, GEM Corporation) (rrr Note 1). 

Previously, the synthesis of peptidic materials was seen as a cumbersome and expensive method that lacked the precision of other polymeric synthesis methods [3, 10, 13, 16, 18, 33, 36-38, 63]. Synthesis of peptides has quickly advanced with cheaper methods, larger yield quantities, greater precision, and more adaptive equipment and methodology. There are two main pathways of creating a large number of customized peptide sequences, (a) engineered/recombinant DNA synthesis or (b) synthetic solid phase peptide synthesis (SPPS) [1,13, 18, 20, 33, 36, 39 5, 64, 65]. These methods are preferred to traditional methods because of the yield, automation, and time saved when compared to manual organic synthesis. 

The synthesis can be carried out manually or automated using a Symphony/Multiplex multiple peptide synthesizer or an Argonaut Nautilus. 

A wide choice of peptide synthesizers is currently available, ranging from manual to fully automated. They are all based on solid-phase peptide synthesis methodologies in which either f-butoxy carbonyl (t-boc) (11), or 9-fluor-enylmethoxycarbonyl (Fmoc) (12) is the major protecting group during synthesis. A detailed description of peptide synthesis is clearly beyond the scope of this chapter, and further information on practical and theoretical approaches to this chemistry may be found elsewhere (13-15). However, a brief outline of solid-phase synthesis may prove useful. 

Solid-phase synthesis of the peptide was carried out manually on a Rink amide resin (435 mg, substitution level 0.46 mmol -g 1) applying the Fmoc strategy. After removal of the Fmoc group attached to the resin with 20% piperidine in DMF (20 min), Fmoc-Tyr(tBu)-OH (230 mg, 0.5 mmol) was introduced with DIC (78 pL, 0.5 mmol) in the presence of HOBt (68 mg, 0.5 mmol) (reaction time 2h). After... 

Manual Stepwise Solid-Phase Peptide Synthesis General Procedure 177 791... 

Peptides were prepared manually by stepwise solid-phase synthesis methodology using conventional N"-Boc chemistry. Briefly, copoly(styrene/l% DVB)-pMeBHA-HCl resin, [100-200 mesh (substitution... 

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