Optimizing Solid-Phase Protein Synthesis Using CPG-2000 and a Nickel-Cleavable SNAC-tag Linker

Solid-phase chemical ligation (SPCL) is a powerful method for simplifying protein synthesis but faces challenges with orthogonal protection strategies, suboptimal solid supports, and limited cleavable linkers. In this study, we optimized SPCL by combining low-loading controlled-pore glass (CPG-2000) with a nickel-cleavable SNAC-tag linker. This system enabled the successful assembly of five peptide fragments and the efficient synthesis of a 131-amino-acid de novo protein, achieving a 25% yield.