Peptide solid-phase, fully automated

W van den Nest, S Yuval, F Albericio. Cu(OBt)2 and Cu(OAt)2, copper(II)-based racemization suppressors ready for use in fully automated solid-phase peptide synthesis. J Pept Sci 7, 115, 2001. 

Poor peptide solubility has been identified as a major obstacle to efficient segment assembly of peptides on a solid phase particularly for fully protected strategies. 1,5,6 Backbone protection directly addresses this problem and is the basis for a general route to the synthesis of small proteins. 7,8 However, the inclusion of backbone protection in a synthesis is not yet automated or routine and requires careful planning. 

Less reactive than acyl halides, but still suitable for difficult couplings, are symmetric or mixed anhydrides (e.g. with pivalic or 2,6-dichlorobenzoic acid) and HOAt-derived active esters. HOBt esters smoothly acylate primary or secondary aliphatic amines, including amino acid esters or amides, without concomitant esterification of alcohols or phenols [34], HOBt esters are the most commonly used type of activated esters in automated solid-phase peptide synthesis. For reasons not yet fully understood, acylations with HOBt esters or halophenyl esters can be effectively catalyzed by HOBt and HOAt [3], and mixtures of BOP (in situ formation of HOBt esters) and HOBt are among the most efficient coupling agents for solid-phase peptide synthesis [2]. In acylations with activated amino acid derivatives, the addition of HOBt or HOAt also retards racemization [4,12,35]. 

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. 

For almost lOyears, automated solid-phase synthesis of oligosaccharides was performed on a modified peptide synthesizer. The first prototype of a specially made oligosaccharide synthesizer [39] features fully automated tailor-made syringe pump systems for accurate delivery of reagents. 

Saneii, H. H. and Shannon, J. D. (1994) Fully automated solid phase synthesis of combinatorial libraries on the peptide librarian. In Innovation and Perspectives in Solid Phase Synthesis (Epton, R., ed.), Intercept, Andover, UK, pp. 335-338. 

A wide choice of peptide-synthesizers is currently available, ranging from manual to fully automated. They are all based on solid-phase peptide syn thesis methodologies in which either fboc (fbutoxy carbonyl) or Fmoc... 

In 1992, ACT replaced the Model 350 with the Model 396 Multiple Biomolecular Synthesizer. This instrument possesses two robotic arms, a variable speed orbital mixer, nitrogen assisted bottom filtration, a built-in ventilation system, protocols for both Fmoc and Boc synthesis and fully automated on-board cleavage for the Fmoc syntheses. It has reactor blocks of 8, 16, 40, and 96 wells for the synthesis of peptides from 5 pmol to 1 mmol. The Model 396 MBS also has a heater/cooler option for applications in solid-phase organic synthesis. 

Cameron, L R, Holder, J L, Meldal, M, Sbeppard, R C, Peptide synthesis. Part 13. Feedback control in solid phase synthesis. Use of fluorenylmethoxycarbonyl amino acid 3,4-dihydro-4-oxo-l,2,3-benzotriazin-3-yl esters in a fully automated system, J. Chem. Soc. Perkin Trans. 1, 2895-2901, 1988. 

To solve the problem, we have developed a highly specific and sensitive assay for C-peptide in serum and plasma using specific monoclonal antibody (MoAb) to the N-terminal of the C-peptide molecule. In this report, we describe the assay performance of C-peptide on the LUMIPULSE system. The system is a fully automated chemiluminescent enzyme immunoassay (CLEIA) system that uses AMPPD as a substrate for alkaline phosphatase and ferrite micro-particles as a solid phase. ... 

Synthesis on solid supports was first developed by Merrifield [1] for the assembly of peptides. It has expanded to include many different applications including oligonucleotide, carbohydrate, and small-molecule assembly (see Chapters 11 and 14). The repetitive cycle of steps involved in the solid-phase synthesis of biopolymers can be performed manually using simple laboratory equipment or fully automated with sophisticated instrumentation. This chapter examines typical solid-phase reaction kinetics to identify factors that can improve the efficiency of both manual and automated synthesis. The hardware and software features of automated solid-phase instruments are also discussed. The focus of this discussion is not on particular commercial model synthesizers but on the basic principles of instrument operation. These considerations can assist in the design, purchase, or use of automated equipment for solid-phase synthesis. Most contrasting features have advantages and disadvantages and the proper choice of instrumentation depends on the synthetic needs of the user. 

Solid-Phase Peptide Synthesis of Decapeptide (H-WFULISTIM-NHi) Derived from the C-Terminus of MuLV CTL Epitope Using a Fully Automated Peptide Synthesizer Syro Wave ... 

Equipment Fully automated X-Y robot using a single-mode microwave-assisted solid-phase peptide synthesizer (Syro Wave , Biotage AB). 

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