Automated solid-phase peptide synthesis

GE Reid, RJ Simpson. Automated solid-phase peptide synthesis use of 2-(17f-ben-zotriazol-l-yl)-l,l,3,3-tetramethyluronium tetrafluoroborate for coupling of tert-butyloxycarbonyl amino acids. Anal Biochem 200, 301, 1992. 

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. 

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]. 

Green fluorescent protein as a marker Section 4.3.5 Immunoelectron microscopy Section 4.3.5 Automated solid-phase peptide synthesis Section 4.4 Nuclear magnetic resonance spectroscopy Section 4.5.1... 

R. Bruce Merrifield was born in 1921 and received a B.S. and a Ph.D. from the University of California, Los Angeles. He is a professor of chemistry at Rockefeller University. Merrifield received the 1984 Nobel Prize in chemistry for developing automated solid-phase peptide synthesis. 

In the Merrifield method, the C-terminal amino acid is covalently attached to a solid support contained in a column. Each N-terminal blocked amino acid is added one at a time, along with other needed reagents, so the protein is synthesized from the C-terminal end to the N-terminal end. Notice that this is opposite to the way proteins are synthesized in nature (from the N-terminal end to the C-terminal end Section 27.13). Because it uses a solid support and is automated, Merrifield s method of protein synthesis is called automated solid-phase peptide synthesis. 

L-amino acid (p. 964) amino acid analyzer (p. 970) amino acid residue (p. 959) anion-exchange resin (p. 970) antiparallel )S-pleated sheet (p. 990) automated solid-phase peptide synthesis (p. 980)... 

First we shall consider reactions for traditional chemical synthesis of peptides and then we look at reactions used in automated solid-phase peptide synthesis. The method for solid-phase peptide synthesis was invented by R. B. Merrifield (Rockefeller University), for which he earned the 1984 Nobel Prize in Chemistry. Solid-phase p>eptide synthesis reactions are so reliable that they have been incorporated into machines called peptide synthesizers (Section 24.7D). 

ROHR" FIGURE 24.8 A method for automated solid-phase peptide synthesis. 

Key words Automated solid-phase peptide synthesis. Microwave heating, Parallel peptide synthesis... 

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