In situ neutralization in boc-chemistry solid phase peptide synthesis: Rapid, high yield assembly of difficult sequences

Schnolzer, Martina, Alewood, Paul, Jones, Alun, Alewood, Dianne and Kent, Stephen B. H. (2007) In situ neutralization in boc-chemistry solid phase peptide synthesis: Rapid, high yield assembly of difficult sequences. International Journal of Peptide Research And Therapeutics, 13 1-2: 31-44. doi:10.1007/s10989-006-9059-7


Author Schnolzer, Martina
Alewood, Paul
Jones, Alun
Alewood, Dianne
Kent, Stephen B. H.
Title In situ neutralization in boc-chemistry solid phase peptide synthesis: Rapid, high yield assembly of difficult sequences
Journal name International Journal of Peptide Research And Therapeutics   Check publisher's open access policy
ISSN 1573-3149
Publication date 2007
Sub-type Article (original research)
DOI 10.1007/s10989-006-9059-7
Volume 13
Issue 1-2
Start page 31
End page 44
Total pages 14
Place of publication New York
Publisher Springer
Collection year 2008
Language eng
Subject C1
0399 Other Chemical Sciences
Abstract Simple, effective protocols have been developed for manual and machine-assisted Boc-chemistry solid phase peptide synthesis on polystyrene resins. These use in situ neutralization [i.e. neutralization simultaneous with coupling], high concentrations (> 0.2 M) of Boc-amino acid-OBt esters plus base for rapid coupling, 100% TFA for rapid Boc group removal, and a single short (30 s) DMF flow wash between deprotection/coupling and between coupling/deprotection. Single 10 min coupling times were used throughout. Overall cycle times were 15 min for manual and 19 min for machine-assisted synthesis (75 residues per day). No racemization was detected in the (.)base-catalyzed coupling step. Several side reactions were studied, and eliminated. These included: pyrrolidonecarboxylic acid formation from Gln in hot TFA-DMF; chain-termination by reaction with excess HBTU; and, chain termination by acetylation (from HOAc in commercial Boc-amino acids). The in situ neutralization protocols gave a significant increase in the efficiency of chain assembly, especially for "difficult" sequences arising from sequence-dependent peptide chain aggregation in standard (neutralization prior to coupling) Boc-chemistry SPPS protocols or in Fmoc-chemistry SPPS. Reported syntheses include HIV-1 protease(1-50,Cys.amide), HIV-1 protease(53-99), and the full length HIV-l protease(1-99).
Keyword Biochemistry & Molecular Biology
difficult sequences
in situ neutralization
ion-spray mass spectrometry
solid phase peptide synthesis (SPPS)
Chemical Synthesis
Hiv Protease
By-products
Proteins
Reagent
Q-Index Code C1
Q-Index Status Confirmed Code

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
2008 Higher Education Research Data Collection
 
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Created: Mon, 18 Feb 2008, 17:29:54 EST