Cysteine(Cys)-specific bioconjugation has widespread applications in the synthesis of protein conjugates,particularly for the functionalization of antibodies.Here,we report the discovery of transstyryl sulfonyl fluori...Cysteine(Cys)-specific bioconjugation has widespread applications in the synthesis of protein conjugates,particularly for the functionalization of antibodies.Here,we report the discovery of transstyryl sulfonyl fluoride(SSF)as a near-perfect Michael acceptor for Cys-specific protein bioconjugation.Compared to maleimides,which are predominantly used,SSF exhibited better chemoselectivity,selfstability,and conjugate stability while maintaining comparable reactivity.Using SSF-derived probes,proteins can be readily modified on the Cys residue(s)to install functionalities,for example,fluorescent dyes,toxins,and oligonucleotides,without influencing the activity.Further applications of SSF-derived serum-stable antibody-drug conjugates and PD-L1 nanobody-oligo conjugates demonstrate the great translational value of SSF-based bioconjugation in drug development and single-cell sequencing.展开更多
基金Financial support from the National Key R&D Program of China(grant no.2019YFA09006600)the National Natural Science Foundation of China(grant nos.21977048 and 92053111)+2 种基金the Natural Science Foundation of Jiangsu Province(grant no.BK20202004)the Beijing National Laboratory for Molecular Sciences(grant no.BNLMS20200)the Jiangsu Specially-Appointed Professor Plan,and the Program for Innovative Talents and Entrepreneur in Jiangsu is gratefully acknowledged.Q.Z.is the Connie and Bob Lurie Fellow of the Damon Runyon Cancer Research Foundation(DRG-2434-21).
文摘Cysteine(Cys)-specific bioconjugation has widespread applications in the synthesis of protein conjugates,particularly for the functionalization of antibodies.Here,we report the discovery of transstyryl sulfonyl fluoride(SSF)as a near-perfect Michael acceptor for Cys-specific protein bioconjugation.Compared to maleimides,which are predominantly used,SSF exhibited better chemoselectivity,selfstability,and conjugate stability while maintaining comparable reactivity.Using SSF-derived probes,proteins can be readily modified on the Cys residue(s)to install functionalities,for example,fluorescent dyes,toxins,and oligonucleotides,without influencing the activity.Further applications of SSF-derived serum-stable antibody-drug conjugates and PD-L1 nanobody-oligo conjugates demonstrate the great translational value of SSF-based bioconjugation in drug development and single-cell sequencing.
