We report a rapid, environment-friendly, and costeffective fi nishing method for cotton textiles by using the isocyanate-coupled sulfopropylbetaine (NCO-sulfopropylbetaine) as the AFA through covalent bond. The sulfopropylbetaine-fi nished cotton textile performs the following functions: (i) resists initial bacterial attachment; (ii) kills attached bacteria; and (iii) releases dead bacteria. Moreover, the hydrophilicity, moisture/air permeability, and mechanical properties of cotton textiles were improved after fi nishing.The antibacterial mechanism of NCO-sulfopropylbetaine was also investigated by isobaric tags for relative and absolute quantitation (iTRAQ) proteomics. Most intriguingly, NCOsulfopropylbetaine shows nontoxicity and good biocompatibility in vivo.