Efficient Site‐Selective Immobilization of Aldehyde‐Tagged Peptides and Proteins by Knoevenagel Ligation

The aldehyde tag is appropriate to selectively label proteins, prepare antibody-drug conjugates or immobilize enzymes antibodies for biotechnological and medical applications. cysteine within the consensus sequence CxPxR of specifically oxidized by formylglycine-generating enzyme (FGE) non-canonical electrophilic amino acid Cα-formylglycine (FGly). Subsequent reductive amination a common method site-directed immobilization, which usually results in poor immobilization efficiency due reaction conditions. Here, we introduce new solid support like agarose modified with an aryl substituted pyrazolone (Knoevenagel reagent) that was obtained facile efficient 2-step synthesis. allowed site-selective aldehyde-containing small molecules, peptides proteins – particular at physiological pH (6.2–8.2) without any additive catalyst needed. In comparison amination, higher loadings activities were achieved various buffers different concentrations temperatures.