Molecularly imprinted polymers(MIPs),as important mimics of antibodies,are chemically synthesized by polymerization in the presence of a target compound.MIPs have found wide applications in important fileds.However,th...Molecularly imprinted polymers(MIPs),as important mimics of antibodies,are chemically synthesized by polymerization in the presence of a target compound.MIPs have found wide applications in important fileds.However,the current molecular imprinting technology suffers from a dilemma;there is often a compromise between the best affinity and the best specificity for MIPs prepared under optimized condi-tions.Herein,we proposed a new strategy called molecular imprinting and cladding(MIC)to solve this issue.The principle is straightforward;after molecular imprinting,a chemically inert cladding thinlayer is generated to precisely cover non-imprinted area.We further proposed a special MIC approach for con-trollably engineering protein binders.The prepared cladded MIPs(cMIPs)exhibited significantly improved affinity and specificity.The general applicability of the proposed strategy and method was ver-ified by engineering of cMIPs for the recognition of a variety of different proteins.The feasibility of cMIPs for real applications was demonstrated by fluorescence imaging of cancer cells against normal cells and immunoassay of C-peptide in human urine.This study opened up a new avenue for controllably engi-neering protein-specific antibody mimics with excellent recognition properties,holding great prospective in important applications such as disease diagnosis and nanomedicine.展开更多
The constantly mutating severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)poses great risk of efficacy loss to the present neutralizing therapeutics.Thus,it is urgently needed to develop versatile strategies ...The constantly mutating severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)poses great risk of efficacy loss to the present neutralizing therapeutics.Thus,it is urgently needed to develop versatile strategies that enable rapid design and engineering of potent neutralizing therapeutics for newly emerging variants.Herein,we present an unprecedented DNA nanocrown that can topologically match and multivalently bind the S-trimer of SARS-CoV-2 and thereby inhibit its infection to host cells.A neutralizing aptamer binding the N-terminal domain(NTD)supersite of the S protein was first screened and identified.It was further elaborately engineered onto the best fitting tetrahedral DNA nanostructure to form a spike protein-capping nanocrown,which can effectively block not only wild-type(WT)SARS-CoV-2 pseudovirus,but also several important mutants including D614G,N501Y,andΔ69–70.Significantly,it can evidently diminish the RNA copies of authentic WT SARS-CoV-2 in host cells by 4.6 orders of magnitude.Therefore,utilizing the aptamer selection method and the dedicated engineering route,our topology-matching DNA framework provides a versatile platform for SARS-CoV-2 inhibition and has the potential to be facilely expanded to newly emerging variants and other fatal coronaviruses.展开更多
基金the Key Program of the National Natural Science Foundation of China(21834003)the National Science Fund for Distinguished Young Scholars(21425520)the Excellent Research Program(ZYJH004)from Nanjing University to ZL。
文摘Molecularly imprinted polymers(MIPs),as important mimics of antibodies,are chemically synthesized by polymerization in the presence of a target compound.MIPs have found wide applications in important fileds.However,the current molecular imprinting technology suffers from a dilemma;there is often a compromise between the best affinity and the best specificity for MIPs prepared under optimized condi-tions.Herein,we proposed a new strategy called molecular imprinting and cladding(MIC)to solve this issue.The principle is straightforward;after molecular imprinting,a chemically inert cladding thinlayer is generated to precisely cover non-imprinted area.We further proposed a special MIC approach for con-trollably engineering protein binders.The prepared cladded MIPs(cMIPs)exhibited significantly improved affinity and specificity.The general applicability of the proposed strategy and method was ver-ified by engineering of cMIPs for the recognition of a variety of different proteins.The feasibility of cMIPs for real applications was demonstrated by fluorescence imaging of cancer cells against normal cells and immunoassay of C-peptide in human urine.This study opened up a new avenue for controllably engi-neering protein-specific antibody mimics with excellent recognition properties,holding great prospective in important applications such as disease diagnosis and nanomedicine.
基金This work was supported by the Key Grant(grant no.21834003)from the National Natural Science Foundation of Chinathe National Key R&D Program of China(grant no.2018YFC0910301)from the Ministry of Science and Technology of Chinathe Excellent Research Program of Nanjing University(grant no.ZYJH004)to Z.L.
文摘The constantly mutating severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)poses great risk of efficacy loss to the present neutralizing therapeutics.Thus,it is urgently needed to develop versatile strategies that enable rapid design and engineering of potent neutralizing therapeutics for newly emerging variants.Herein,we present an unprecedented DNA nanocrown that can topologically match and multivalently bind the S-trimer of SARS-CoV-2 and thereby inhibit its infection to host cells.A neutralizing aptamer binding the N-terminal domain(NTD)supersite of the S protein was first screened and identified.It was further elaborately engineered onto the best fitting tetrahedral DNA nanostructure to form a spike protein-capping nanocrown,which can effectively block not only wild-type(WT)SARS-CoV-2 pseudovirus,but also several important mutants including D614G,N501Y,andΔ69–70.Significantly,it can evidently diminish the RNA copies of authentic WT SARS-CoV-2 in host cells by 4.6 orders of magnitude.Therefore,utilizing the aptamer selection method and the dedicated engineering route,our topology-matching DNA framework provides a versatile platform for SARS-CoV-2 inhibition and has the potential to be facilely expanded to newly emerging variants and other fatal coronaviruses.
