Precise Synthesis of Proteins/Peptides: Advances in Single-Site Selective Chemical Modification and Automated Synthesis Technologies★

doi:10.6023/cjoc202505023

Abstract

The precise synthesis of proteins and peptides is fundamental for developing innovative therapeutics, high-per- formance molecular probes, and advanced functional materials. Recent advances in single-site selective chemical modification and automated synthesis technologies have significantly improved precision in sequence control, enabled site-specific introduction of functional groups and diversified modification. These developments have effectively addressed the issue of product heterogeneity from traditional non-selective modifications while substantially improving the efficiency and precision of complex sequence synthesis. This review summarizes the research progress in single-site modification strategies for proteins/peptides over the past five years (2020~2025), focusing on site-specific direct modification, ligand-directed modification, and linchpin-directed modification. It also provides a systematic evaluation of recent developments in automated precision synthesis technologies. These technological innovations have not only enhanced the capability for controllable synthesis and precise functionalization of protein/peptide, but also created new opportunities in drug development, molecular probe engineering, and bioconjugation applications. Looking ahead, the development and application of novel bioorthogonal reagents, the integration of innovative dynamic modification technologies, and AI-assisted synthesis route optimization will emerge as key research directions, promising to drive breakthroughs in protein precision synthesis.

Key words: protein, peptide, precise chemical synthesis, single-site modification, automated synthesis

Cite this article

Cheng Ren, Chengxi Li. Precise Synthesis of Proteins/Peptides: Advances in Single-Site Selective Chemical Modification and Automated Synthesis Technologies^★[J]. Chinese Journal of Organic Chemistry, 2025, 45(9): 3128-3147.

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Fig. & Tab. 19

Figure 1. Single-site direct chemical modification of proteins/peptides

Figure 2. Single-site selective modification of the N-terminal amino group in proteins/peptides

Figure 3. Single-site selective modification of N-terminal backbone amide bonds in proteins/peptides

Figure 4. Single-site selective modification of N-terminal proline in proteins/peptides

Figure 5. Single-site selective modification of N-terminal glycine in proteins/peptides

Figure 6. Single-site selective modification of N-terminal cysteine in proteins/peptides

Figure 7. Protein/peptide C-terminal single-site selective modification

Figure 8. Ligand-directed single-site modification

Figure 9. Ligand-directed lysine single-site selective modification

Figure 10. CoLDR strategy for cysteine single-site selective modification

Figure 11. Single-site selective modification based on affinity-guided catalytic strategy

Figure 12. Linchpin-directed modification

Figure 13. Single-site selective modification of lysine residue by FK1-spacer-FK2

Figure 14. Single-site selective modification of lysine residue by FC1-spacer-FK2

Figure 15. Linchpin-directed single-site selective modification of aspartate or histidine residue

Figure 16. Automated flow synthesis technology for peptides

Figure 17. Automated parallel synthesis technology for peptides

Figure 18. Automated flow synthesis technology for proteins

Figure 19. Automated precise synthesis of peptides guided by machine learning

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