Dual Plug-and-Display Synthetic Assembly Using Orthogonal Reactive Proteins for Twin Antigen Immunization

Abstract

Engineering modular platforms to control biomolecular architecture can advance both the understanding and the manipulation of biological systems. Icosahedral particles uniformly displaying single antigens stimulate potent immune activation and have been successful in various licensed vaccines. However, it remains challenging to display multiple antigens on a single particle and to induce broader immunity protective across strains or even against distinct diseases. Here, we design a dually addressable synthetic nanoparticle by engineering the multimerizing coiled-coil IMX313 and two orthogonally reactive split proteins. SpyCatcher protein forms an isopeptide bond with SpyTag peptide through spontaneous amidation. SnoopCatcher forms an isopeptide bond with SnoopTag peptide through transamidation. SpyCatcher-IMX-SnoopCatcher provides a modular platform, whereby SpyTag-antigen and SnoopTag-antigen can be multimerized on opposite faces of the particle simply upon mixing. We demonstrate efficient derivatization of the platform with model proteins and complex pathogen-derived antigens. SpyCatcher-IMX-SnoopCatcher was expressed in Escherichia coli and was resilient to lyophilization or extreme temperatures. For the next generation of malaria vaccines, blocking the transmission of the parasite from human to mosquito is an important goal. SpyCatcher-IMX-SnoopCatcher multimerization of the leading transmission-blocking antigens Pfs25 and Pfs28 greatly enhanced the antibody response to both antigens in comparison to the monomeric proteins. This dual plug-and-display architecture should help to accelerate vaccine development for malaria and other diseases.

Development of target‐tunable P22 VLP‐based delivery nanoplatforms using bacterial superglue

“…We also demonstrated that those conjugates exhibited a range of different functions (Bae et al, ; Choi et al, ; Moon et al, ). The ST/SC protein ligation system has been applied for displaying various antigenic proteins and targeting ligands on VLPs and their effective immune response inductions and cell‐specific gene delivery were successfully demonstrated (Brune et al, , ; Bruun, Andersson, Draper, & Howarth, ; Janitzek et al, ; Kasa, Raneni, Chamoun‐Emanuelli, Wright, & Chen, ; Ke et al, ; Leneghan et al, ; Singh et al, ; Thrane et al, ).…”

Designing Multivalent Ligands to Control Biological Interactions: From Vaccines and Cellular Effectors to Targeted Drug Delivery

“…The Howarth group has developed orthogonally reactive split proteins such as SpyCatcher/SpyTag—by splitting a domain from the fibronectin‐binding protein of Streptococcus pyrogenes —and SnoopCatcher/SnoopTag—by splitting an adhesin from Streptococcus pneumonia . More recently, Brune et al . genetically fused the sequences for SpyCatcher and SnoopCatcher to different termini of the multimerized coiled‐coil IMX313 scaffold and expressed the final construct in E. coli (Figure ).…”

“…Due to strong and rapid reactions between the tags and the complementary Catchers, the authors were able to covalently link the antigens to the IMX platform, which was demonstrated to be stable even under non‐ideal conditions. In addition, during animal studies, the multivalent construct resulted in an antibody response 10‐fold higher than the one using soluble monovalent antigens …”

“…The final construct is a nanoparticle displaying multiple copies of the two different antigens. Figure reprinted from with permission. Copyright 2017 American Chemical Society.…”

“…The Howarth group has developed orthogonally reactive split proteins such as SpyCatcher/SpyTag-bys plitting a domain from the fibronectin-binding protein of Streptococcus pyrogenes-and SnoopCatcher/SnoopTag-by splitting an adhesin from Streptococcus pneumonia. [18] More recently,B rune et al [19] genetically fused the sequences for SpyCatcher and SnoopCatcher to different termini of the multimerized coiledcoil IMX313 scaffold and expressed the final construct in E. coli ( Figure 1). In parallel, two malarial antigens-Pfs25 and Pfs28-were fused to the peptides SpyTag and SnoopTag, respectively.D ue to stronga nd rapid reactions between the tags and the complementary Catchers, the authors were able to covalentlyl ink the antigenst ot he IMX platform, whichw as demonstrated to be stable even under non-ideal conditions.…”