Serine-mediated hydrazone ligation displaying insulin-like peptides on M13 phage pIII

Phage display has emerged as a tool for the discovery of therapeutic antibodies and proteins. However, the effective display and engineering of structurally complex proteins, such as insulin, pose significant challenges due to the sequence of insulin, which is composed of two peptide chains linked b...

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Bibliographic Details
Published in:Organic & biomolecular chemistry 2023-11, Vol.21 (44), p.892-899
Main Authors: Zhang, Yi Wolf, Zheng, Nan, Chou, Danny Hung-Chieh
Format: Article
Language:English
Subjects:
Antibodies
Diabetes mellitus
Disulfide bonds
Hydrazones
Insulin
Peptides
Phage display
Phages
Proteins
Serine
Trypsin
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Summary:Phage display has emerged as a tool for the discovery of therapeutic antibodies and proteins. However, the effective display and engineering of structurally complex proteins, such as insulin, pose significant challenges due to the sequence of insulin, which is composed of two peptide chains linked by three disulfide bonds. In this study, we developed a new approach for the display of insulin-like peptides on M13 phage pIII, employing N-terminal serine-mediated hydrazone ligation. The insulin-displaying phage retains the biological binding affinity of human insulin. To address the viability loss after ligation, we introduced a trypsin-cleavable spacer on pIII, enabling insulin-displayed phage library selection. This method offers a general pathway for the display of structurally complex proteins on pIII, enhancing the practicality of selecting chemically modified phage libraries and opening avenues for the engineering of new insulin analogs for the treatment of diabetes by using phage display. An new method displays insulin-like peptides on M13 phage via hydrazide-Fmoc-small insulin and serine-hydrazone ligation. This approach addresses insulin's structural challenges and highlights potential for displaying complex proteins on phage.
ISSN:1477-0520
1477-0539
DOI:10.1039/d3ob01487h