Document text
Principal Investigator: Patrick Duffy
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2021
Award: $780,824
Funding agency: National Institute of Allergy and Infectious Diseases
LMIV produced and obtained antigens for multiple COVID-19 serology assays. First, plasmids were obtained for production of full-length spike and RBD used in the assay developed by Florian Krammer at Mount Sinai Hospital in New York. After 3 successful production runs of COVID-19 spike proteins used in the Krammer assay, the in-house proteins were shown to perform in a manner identical to proteins produced in the Krammer group. Second, proteins were produced and obtained for a serological assay developed by Kaitlyn Sadtler (NIBIB). LMIV initiated a collaboration with Dominic Esposito (NCI) and obtained enough protein for the entire serological survey program in Africa. LMIV also obtained the plasmids for both proteins used in the Sadtler assay and produced at high yields the RBD protein to be used for in house assay development. Third, LMIV designed a construct expressing the full-length nucleocapsid protein in mammalian cells and produced full-length nucleocapsid at high levels to use in serology assay to compare to spike and RBD. Fourth, LMIV constructed a plasmid and expressed the Ace2 binding domain and demonstrated binding of RBD to Ace2 in an ELISA based format establishing an assay at LMIV that can be used to evaluate antibodies that block Ace2 binding to RBD and spike. Additionally, the first COVID-19 animal immunization was performed at LMIV and this study demonstrated immunogenicity of spike and RBD in mice instituting a benchmark for LMIV going forward for new immunogen design.
SARS-CoV-2 antigens (full-length Spike (S) and Receptor binding domain (RBD)) from commercial sources (Ray Biotech, US) and academic partner laboratories (Vaccine Research Center, NIH; Icahn School of Medicine at Mt Sinai; Ragon institute, MGH, MIT)) were compared using either the assay established by the Krammer laboratory (Mt Sinai) or the Sadtler laboratory (NIBIB, NIH), as per their defined standard operating procedures. In addition, antibodies against the Nucleocapsid (N) protein (antigen produced at LMIV) were measured using the Sadtler assay.
US COVID nave (n =20), acute convalescent sera samples from US COVID19 patients (n=114) and Mali sera from three active cohorts (Bancoumana, Ouelessebougou and Kalifabougou, n = 248) were screened (in duplicates) for reactivity to COVID-19 antigens. In addition, the Malian sera were also screened for antibodies against Spike proteins from other known betacoronaviruses (SARS, MERS, OC43 and HKU1) kindly provided by Kaitlyn Sadtler.
In July 2020, we commenced a community COVID-19 seroprevalence study at existing clinical trials sites in Mali. This study is a Public Health Surveillance Activity in collaboration with the Ministry of Health in Mali to describe the sero-epidemiology of COVID-19 in urban and rural populations. Using the demographic and clinical information collected from participants, we will describe the age-stratified seroprevalence and fraction of asymptomatic/pauci-symptomatic cases to help understand the penetration of SARS-CoV-2 into the community.
In parallel to the serosurvey program described above, LMIV has initiated a project on developing vaccine candidates against SARS-CoV-2 to address the pandemic caused by this virus. As part of this parallel program, we used antigens generated in the Serosurvey program as benchmark immunogens to pilot small animal immunization studies and platforms. LMIV has over the years developed a Conjugate Nanoparticle Vaccine technology for malaria using a synthetic approach that generates a nanoparticle structure with antigen and a carrier protein are crosslinked at an approximately 1:1 ratio. A malaria transmission blocking vaccine employing this technology is currently undergoing Phase II clinical trial in malaria endemic areas in Mali. This technology has proven effective in enhancing the immunogenicity of poorly immunogenic subunit protein antigens. We have adapted this technology to develop a second-generation vaccine against SARS-CoV-2 using the Receptor Binding Domain (RBD) of the Spike protein as the antigen. RDB is not a highly immunogenic protein, but conjugating this to a carrier protein, we believe, will make the conjugate an effective vaccine candidate. By using RBD as antigen, we hope to focus the immune response to the receptor binding interface and generate potent neutralizing monoclonal antibody that blocks the virus binding to ACE2 receptor. Thus far we have synthesized and characterized a number of chemically crosslinked conjugates of two different RBD constructs to a carrier protein EcoCRM. This carrier is shown to be an effective carrier in our malaria vaccine studies. We are preparing to evaluate the immunogenicity of these conjugates compared to unconjugated RBD monomers, in mouse immunogenicity studies in combination with two different adjuvants.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><2019-nCoV vaccine><ACE2><Acute><Address><Adjuvant><Africa><Age><Animals><Antibodies><Antigens><Area><Assay><Benchmarking><Best Practice Analysis><Binding><Bioassay><Biologic Assays><Biological Assay><Biotech><Biotechnology><Blocking Antibodies><COVID><COVID crisis><COVID epidemic><COVID infected patient><COVID pandemic><COVID patient><COVID positive patient><COVID-19><COVID-19 S protein><COVID-19 antigen><COVID-19 crisis><COVID-19 epidemic><COVID-19 global health crisis><COVID-19 global pandemic><COVID-19 health crisis><COVID-19 infected patient><COVID-19 pandemic><COVID-19 patient><COVID-19 positive patient><COVID-19 public health crisis><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 vaccine><COVID-19 virus><COVID19><COVID19 S protein><COVID19 crisis><COVID19 epidemic><COVID19 global health crisis><COVID19 global pandemic><COVID19 health crisis><COVID19 pandemic><COVID19 patient><COVID19 positive patient><COVID19 public health crisis><COVID19 spike glycoprotein><COVID19 spike protein><COVID19 vaccine><COVID19 virus><CV-19><CV19><Carrier Proteins><Chemicals><Clinical><Clinical Trials><CoV disease><CoV-2><CoV2><Collaborations><Communities><ELISA><Enzyme-Linked Immunosorbent Assay><Epidemiology><Generations><Goals><Health><Hospitals><Household><Immune response><Immunization><Immunologic Sensitization><Immunologic Stimulation><Immunological Sensitization><Immunological Stimulation><Immunological response><Immunostimulation><Individual><Institutes><Laboratories><Length><MERS><MERS coronavirus disease><MERS-CoV disease><Malaria><Malaria Vaccines><Malarial Vaccines><Mali><Mammalian Cell><Measures><Mice><Mice Mammals><Middle East Respiratory Syndrome><Middle East Respiratory Syndrome CoV disease><Middle East Respiratory Syndrome coronavirus disease><Middle Eastern Respiratory Syndrome><Middle Eastern Respiratory Syndrome CoV disease><Middle Eastern Respiratory Syndrome coronavirus disease><Molecular Interaction><Murine><Mus><NIBIB><NIH><National Institute of Biomedical Imaging and Bioengineering><National Institutes of Health><New York><Nucleocapsid><Nucleocapsid Proteins><Outcome><Paludism><Participant><Penetration><Phase><Phase 2 Clinical Trials><Phase II Clinical Trials><Plasmids><Plasmodium Infections><Population Surveillance><Post-partum Women><Postpartum Women><Pregnant Women><Preparation><Prevalence><Procedures><Production><Protein Binding Domain><Protein Binding Motif><Protein Subunits><Protein-Protein Interaction Domain><Proteins><Public Health Surveillance><Reagent><Receptor Protein><Recombinants><Running><Rural><Rural Population><SARS><SARS corona virus 2><SARS coronavirus disease><SARS-CoV disease><SARS-CoV-2><SARS-CoV-2 S protein><SARS-CoV-2 antigen><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 infected patient><SARS-CoV-2 pandemic><SARS-CoV-2 patient><SARS-CoV-2 positive patient><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV-2 vaccine><SARS-CoV2><SARS-CoV2 S protein><SARS-CoV2 antigen><SARS-CoV2 epidemic><SARS-CoV2 pandemic><SARS-CoV2 spike glycoprotein><SARS-CoV2 spike protein><SARS-CoV2 vaccine><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-coronavirus-2 epidemic><SARS-coronavirus-2 pandemic><SARS-coronavirus-2 vaccine><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Sampling><Sentinel><Serology test><Seroprevalences><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome CoV 2 epidemic><Severe Acute Respiratory Syndrome CoV 2 pandemic><Severe Acute Respiratory Syndrome CoV 2 vaccine><Severe Acute Respiratory Syndrome CoV disease><Severe Acute Respiratory Syndrome coronavirus disease><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 S protein><Severe acute respiratory syndrome coronavirus 2 epidemic><Severe acute respiratory syndrome coronavirus 2 pandemic><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome coronavirus 2 vaccine><Severe acute respiratory syndrome related corona virus 2><Site><Source><Structure><Technology><Transport Protein Gene><Transport Proteins><Transporter Protein><United States National Institutes of Health><Urban Population><Vaccine Research><Vaccines><Virus><Wuhan coronavirus><ages><angiotensin converting enzyme 2><angiotensin converting enzyme II><assay development><base><beta CoV><beta coronavirus><betaCoV><betacoronavirus><cohort><corona virus disease><corona virus disease 2019><corona virus disease 2019 epidemic><corona virus disease 2019 pandemic><corona virus disease 2019 vaccine><coronavirus disease><coronavirus disease 2019><coronavirus disease 2019 S protein><coronavirus disease 2019 antigen><coronavirus disease 2019 crisis><coronavirus disease 2019 epidemic><coronavirus disease 2019 global health crisis><coronavirus disease 2019 global pandemic><coronavirus disease 2019 health crisis><coronavirus disease 2019 infected patient><coronavirus disease 2019 pandemic><coronavirus disease 2019 patient><coronavirus disease 2019 positive patient><coronavirus disease 2019 public health crisis><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 vaccine><coronavirus disease 2019 virus><coronavirus disease crisis><coronavirus disease epidemic><coronavirus disease infected patient><coronavirus disease pandemic><coronavirus disease patient><coronavirus disease positive patient><coronavirus patient><cross-link><crosslink><design><designing><epidemiologic><epidemiological><expectant mother><expecting mother><hCoV19><host response><immune system response><immunogen><immunogenic><immunogenicity><immunoresponse><malaria transmission-blocking vaccine><medical college><medical schools><monomer><nCoV2><nano particle><nano-sized particle><nanoparticle><nanosized particle><neutralizing mAb><neutralizing monoclonal antibodies><pandemic><pandemic disease><patient infected with COVID><patient infected with COVID-19><patient infected with SARS-CoV-2><patient infected with coronavirus disease><patient infected with coronavirus disease 2019><patient infected with severe acute respiratory syndrome coronavirus 2><patient with COVID><patient with COVID-19><patient with COVID19><patient with SARS-CoV-2><patient with coronavirus disease><patient with coronavirus disease 2019><patient with severe acute respiratory distress syndrome coronavirus 2><performance tests><phase II protocol><pregnant mothers><programs><protein crosslink><receptor><receptor binding><receptor bound><response><school of medicine><serology assay><serology surveillance><serology survey><seropositive><serosurveillance><serosurvey><severe acute respiratory syndrome coronavirus 2 antigen><severe acute respiratory syndrome coronavirus 2 global health crisis><severe acute respiratory syndrome coronavirus 2 global pandemic><severe acute respiratory syndrome coronavirus 2 infected patient><severe acute respiratory syndrome coronavirus 2 patient><severe acute respiratory syndrome coronavirus 2 positive patient><vaccination study><vaccination trial><vaccine against 2019-nCov><vaccine against SARS-CoV-2><vaccine against SARS-CoV2><vaccine against SARS-coronavirus-2><vaccine against Severe Acute Respiratory Syndrome CoV 2><vaccine against Severe acute respiratory syndrome coronavirus 2><vaccine candidate><vaccine for novel coronavirus><vaccine study><vaccine trial><vaccines against malaria><β CoV><β coronavirus><βCoV>