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Thermo Fisher Scientific DEFORMYLATION OF PEPTIDES SYNTHESIZED USING TRP Owner's manual

  • Hello! I've reviewed the User Bulletin regarding the deformylation of peptides synthesized using TRP (CHO) protection from Applied Biosystems. This document details a specific procedure involving dissolving peptides and performing UV analysis, and it provides key information on expected readings. I am ready to answer your questions related to this document and the outlined peptide synthesis process.
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    What is the first step in the deformylation procedure?
    How should a UV spectral analysis look for a peptide containing Trp (CHO)?
User Bulletin No. 18
Peptide Synthesizer
April 28, 1987 (updated 09/2002)
SUBJECT: DEFORMYLATION OF PEPTIDES SYNTHESIZED USING TRP
(CHO) PROTECTION
Author: Alan Culwell and James T. Sparrow
Introduction The formyl (CHO) protecting group of tryptophan is an HF, TFMSA stable species.
Deformylation of a peptide synthesized by using TRP (CHO) requires a separate step
after strong acid cleavage. Since the deformylation of Trp (CHO) can pose problems to
even the experienced peptide chemist, the following procedure should be strictly
followed.
Procedure 1. Dissolve the peptide containing Trp (CHO) in 6M guanidine HC1 to produce a
concentration between 1 and 10 mg/mL.
2. Perform a UV spectral analysis on the dissolved peptide. For a peptide containing
Trp (CHO), the 300 nm absorbance is greater than the 280 nm absorbance, as
shown on Figure 1.
Figure 1. UV spectral analysis of a peptide containing Trp (CHO) before deformylation with
ethanolamine.
© Copyright 2002, Applied Biosystems
For Research Use Only. Not for use in diagnostic procedures.
Stock No. 118UB03-02
Figure 2. HPLC Profiles of the HF Crude Peptide Gastrin-1.
(A) Before deformylation of the peptide containing Trp (CHO).
(B) After deformylation of the peptide containing Trp (CHO).
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