Method of Monitoring the Number of Amide Bonds in Peptides Using Near-Infrared Spectroscopy

Using near-infrared (NIR) spectroscopy, we aimed to develop a method of monitoring the increasing number of amide bonds with the elongation of the chain length of peptides. Because peptide synthesis can be monitored by evaluating the increasing number of amide bonds with dehydration occurring betwee...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2021-02, Vol.93 (5), p.2758-2766
Main Authors: Ishigaki, Mika, Ito, Atsushi, Hara, Risa, Miyazaki, Shun-ichi, Murayama, Kodai, Yoshikiyo, Keisuke, Yamamoto, Tatsuyuki, Ozaki, Yukihiro
Format: Article
Language:English
Subjects:
Amino acids
Banded structure
Bonding
Calibration
Chains
Chemistry
Dehydration
Elongation
Glycine
I.R. radiation
Infrared spectra
Infrared spectroscopy
Monitoring
Near infrared radiation
Peptide synthesis
Peptides
Polyamide resins
Polyamides
Polypeptides
Protecting groups
Regression analysis
Spectrum analysis
Technology assessment
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Summary:Using near-infrared (NIR) spectroscopy, we aimed to develop a method of monitoring the increasing number of amide bonds with the elongation of the chain length of peptides. Because peptide synthesis can be monitored by evaluating the increasing number of amide bonds with dehydration occurring between amino acids, polyglycine, which has the simplest structure among polyamino acids, was studied, and the key bands whose absorption intensities increased with the elongation of the chain length, such as the bands attributed to glycine, diglycine, triglycine, and tetraglycine, were searched. The bands due to the combinations of the amide A and amide II/III modes in the region of 5000–4500 cm–1 were revealed to be good candidates for key bands, their second derivative intensities increased as the number of amide bonds increased, regardless of pH, solvent species, and the presence of protecting groups. The number of amide bonds was evaluated by a partial least square regression using the abovementioned combination bands, and a calibration model with a high determination coefficient (≥0.99) was constructed. These results not only have demonstrated the usefulness of NIR spectroscopy as a process analytical technology tool for the process of synthesizing the peptide in a microflow reactor but also have provided basic knowledge for analyzing amide bonds in the NIR spectra of proteins, polyamino acids, polypeptides, and polyamides.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.0c03424