Assembly of Immunogenic Protein Particles toward Advanced Synthetic Vaccines.

Immunogenic carrier proteins such as the non-toxic diphtheria toxin variant, cross-reacting material 197 (CRM197), are widely used in subunit vaccine formulations to boost immunogenicity of chemically conjugated antigens. Conjugate vaccines are inherently expensive due to laborious manufacturing steps. Here, this work develops a particulate vaccine platform based on using engineered Escherichia coli to assemble CRM197-antigen fusion proteins into discrete submicron-sized particles. This approach enables precise loading of diverse antigens and epitopes enhancing their immunogenicity. A cost-effective, high-yield, and scalable biomanufacturing process is developed. Purified particulate CRM197-antigen vaccines are ambient-temperature stable. CRM197 particles incorporating pathogen-specific antigens or epitopes from SARS-CoV-2, Streptococcus pyogenes (group A), and Mycobacterium tuberculosis induced cell-mediated and humoral immune responses mediating protective immunity in respective animal models of infection. The CRM197 particle vaccine platform is versatile, enabling co-delivery of selected antigens/epitopes together with immunogenic CRM197 as discrete stable particles avoiding laborious manufacture of soluble CRM197 and antigen followed by chemical conjugation.

[1]  Xing Wu,et al.  Research progress on vaccine efficacy against SARS-CoV-2 variants of concern , 2022, Human vaccines & immunotherapeutics.

[2]  Y. Kawaoka,et al.  Ambient Temperature Stable, Scalable COVID‐19 Polymer Particle Vaccines Induce Protective Immunity , 2021, Advanced healthcare materials.

[3]  S. A. Castro,et al.  A brief review on Group A Streptococcus pathogenesis and vaccine development , 2021, Royal Society Open Science.

[4]  Manisha Pandey,et al.  Preclinical safety and immunogenicity of Streptococcus pyogenes (Strep A) peptide vaccines , 2021, Scientific reports.

[5]  Shuxiong Chen,et al.  Polymeric nanoparticle vaccines to combat emerging and pandemic threats , 2020, Biomaterials.

[6]  Fumihiro Kato,et al.  Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells , 2020, Proceedings of the National Academy of Sciences.

[7]  J. Mascola,et al.  Preclinical Development of a Fusion Peptide Conjugate as an HIV Vaccine Immunogen , 2020, Scientific Reports.

[8]  S. Harding,et al.  Glycoconjugate vaccines: some observations on carrier and production methods. , 2019, Biotechnology & genetic engineering reviews.

[9]  W. M. Hussein,et al.  Recent Advances in the Development of Peptide Vaccines and Their Delivery Systems against Group A Streptococcus , 2019, Vaccines.

[10]  Yu Qin,et al.  Co-delivery of antigen and dual agonists by programmed mannose-targeted cationic lipid-hybrid polymersomes for enhanced vaccination. , 2019, Biomaterials.

[11]  Peifang Wang,et al.  Nanoparticle tracking analysis versus dynamic light scattering: Case study on the effect of Ca2+ and alginate on the aggregation of cerium oxide nanoparticles. , 2018, Journal of hazardous materials.

[12]  Shuxiong Chen,et al.  Design of Bacterial Inclusion Bodies as Antigen Carrier Systems , 2018, Advanced Biosystems.

[13]  Bong-Seong Koo,et al.  Efficient recovery of recombinant CRM197 expressed as inclusion bodies in E.coli , 2018, PloS one.

[14]  K. Kaur,et al.  Analytical Comparability Assessments of 5 Recombinant CRM197 Proteins From Different Manufacturers and Expression Systems. , 2018, Journal of pharmaceutical sciences.

[15]  M. Flórido,et al.  PLGA particulate subunit tuberculosis vaccines promote humoral and Th17 responses but do not enhance control of Mycobacterium tuberculosis infection , 2018, PloS one.

[16]  T. Kleffmann,et al.  Bioengineered polyester beads co-displaying protein and carbohydrate-based antigens induce protective immunity against bacterial infection , 2018, Scientific Reports.

[17]  A. Esser‐Kahn,et al.  Toll-like Receptor Agonist Conjugation: A Chemical Perspective. , 2018, Bioconjugate chemistry.

[18]  B. Rehm.  Bioengineering towards self-assembly of particulate vaccines. , 2017, Current opinion in biotechnology.

[19]  Manisha Pandey,et al.  Enhancing Vaccine Efficacy by Engineering a Complex Synthetic Peptide To Become a Super Immunogen , 2017, The Journal of Immunology.

[20]  R. A. van den Berg,et al.  Cellular and molecular synergy in AS01-adjuvanted vaccines results in an early IFNγ response promoting vaccine immunogenicity , 2017 .

[21]  L. Pirofski,et al.  A semisynthetic Streptococcus pneumoniae serotype 8 glycoconjugate vaccine , 2017, Science Translational Medicine.

[22]  J. Moon,et al.  Designer vaccine nanodiscs for personalized cancer immunotherapy , 2016, Nature materials.

[23]  P. Burkhard,et al.  Vaccine technologies: From whole organisms to rationally designed protein assemblies , 2016, Biochemical Pharmacology.

[24]  Manisha Pandey,et al.  Combinatorial Synthetic Peptide Vaccine Strategy Protects against Hypervirulent CovR/S Mutant Streptococci , 2016, The Journal of Immunology.

[25]  Lyn H Jones,et al.  Recent advances in the molecular design of synthetic vaccines. , 2015, Nature chemistry.

[26]  T. Scriba,et al.  T cells and adaptive immunity to Mycobacterium tuberculosis in humans , 2015, Immunological reviews.

[27]  J. Sterne,et al.  Protection by BCG vaccine against tuberculosis: a systematic review of randomized controlled trials. , 2014, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[28]  Gregory L. Szeto,et al.  Structure-based programming of lymph-node targeting in molecular vaccines , 2014, Nature.

[29]  M. Pichichero.  Protein carriers of conjugate vaccines , 2013, Human vaccines & immunotherapeutics.

[30]  J. Haensler,et al.  An update on safety and immunogenicity of vaccines containing emulsion-based adjuvants , 2013, Expert review of vaccines.

[31]  I. Toth,et al.  Modern Subunit Vaccines: Development, Components, and Research Opportunities , 2013, ChemMedChem.

[32]  G. Schoolnik,et al.  A multistage tuberculosis vaccine that confers efficient protection before and after exposure , 2011, Nature Medicine.

[33]  H. Shinefield.  Overview of the development and current use of CRM(197) conjugate vaccines for pediatric use. , 2010, Vaccine.

[34]  M. Roederer,et al.  T-cell quality in memory and protection: implications for vaccine design , 2008, Nature Reviews Immunology.

[35]  D. Kelly,et al.  CRM197-conjugated serogroup C meningococcal capsular polysaccharide, but not the native polysaccharide, induces persistent antigen-specific memory B cells. , 2006, Blood.

[36]  T. Bártfai,et al.  Protective immunogenicity of two synthetic peptides selected from the amino acid sequence of Bordetella pertussis toxin subunit S1. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[37]  R. Rappuoli,et al.  The amino-acid sequence of two non-toxic mutants of diphtheria toxin: CRM45 and CRM197. , 1984, Nucleic acids research.

[38]  E. N. Fox,et al.  Protective studies with a group A streptococcal M protein vaccine. II. Challange of volenteers after local immunization in the upper respiratory tract. , 1975, The Journal of infectious diseases.