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

Karl D Brune,Sumi Biswas,Iona J Taylor,Florian Brod,Yuanyuan Li,Mark Howarth,Can M Buldun

doi:10.1021/acs.bioconjchem.7b00174

Karl D Brune, Sumi Biswas + Show 5 more

Open Access

https://doi.org/10.1021/acs.bioconjchem.7b00174

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Journal: Bioconjugate chemistry	Publication Date: May 5, 2017
Citations: 87	License type: cc-by

Affiliation: University of Oxford

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.

Highlights

Biological behavior can be exquisitely sensitive to nanoscale molecular organization.[1,2] Understanding and controlling such biomolecular organization is an important goal for basic biological insight[3] and for the generation of novel vaccines
Termini of IMX313 are more closely packed than the N-termini, a longer spacer was added for SnoopCatcher than SpyCatcher to provide steric accessibility for coupling of multiple protein antigens (Figure 1)
SpyCatcher−IMX−SnoopCatcher was expressed most efficiently and with the best purity in E. coli by refolding from inclusion bodies. (We name all constructs with the N-terminal component first and the C-terminal component last.) Particles were purified by Ni−NTA affinity chromatography (Figure 2a)

Summary

Introduction

Biological behavior (for example, stimulation of an immune response) can be exquisitely sensitive to nanoscale molecular organization.[1,2] Understanding and controlling such biomolecular organization is an important goal for basic biological insight[3] and for the generation of novel vaccines. Vaccines were generally based on whole pathogens (attenuated or dead). A notable example is the IMX313 scaffold, engineered from the heptamerizing coiledcoil domain of complement C4 binding protein.[9] IMX313 conjugates have already moved into clinical development.[10,11]

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