Thermophilic Filamentous Fungus C1-Cell-Cloned SARS-CoV-2-Spike-RBD-Subunit-Vaccine Adjuvanted with Aldydrogel®85 Protects K18-hACE2 Mice against Lethal Virus Challenge

Ram Nechooshtan; Sharon Ehrlich; Marika Vitikainen; Arik Makovitzki; Eyal Dor; Hadar Marcus; Idan Hefetz; Shani Pitel; Marilyn Wiebe; Anne Huuskonen; Lilach Cherry; Edith Lupu; Yehuda Sapir; Tzvi Holtzman; Moshe Aftalion; David Gur; Hadas Tamir; Yfat Yahalom-Ronen; Yuval Ramot; Noam Kronfeld; David Zarling; Anne Vallerga; Ronen Tchelet; Abraham Nyska; Markku Saloheimo; Mark Emalfarb; Yakir Ophir

doi:10.3390/vaccines10122119

Thermophilic Filamentous Fungus C1-Cell-Cloned SARS-CoV-2-Spike-RBD-Subunit-Vaccine Adjuvanted with Aldydrogel^®85 Protects K18-hACE2 Mice against Lethal Virus Challenge

Ram Nechooshtan
, Sharon Ehrlich
, Marika Vitikainen
, Arik Makovitzki
, Eyal Dor
, Hadar Marcus
, Idan Hefetz
, Shani Pitel
, Marilyn Wiebe
, Anne Huuskonen
, Lilach Cherry
, Edith Lupu
, Yehuda Sapir
, Tzvi Holtzman
, Moshe Aftalion
, David Gur
, Hadas Tamir
, Yfat Yahalom-Ronen
, Yuval Ramot
, Noam Kronfeld

David Zarling, Anne Vallerga, Ronen Tchelet, Abraham Nyska, Markku Saloheimo, Mark Emalfarb, Yakir Ophir^*

^*Corresponding author for this work

Bioprocess engineering

Israel Institute for Biological Research
Hebrew University of Jerusalem
Hadassah University Medical Centre
Envigo CRS Israel Ltd
Dyadic International Inc.
Dyadic Netherland B.V.
Tel Aviv University
University of California, Davis

Research output: Contribution to journal › Article › Scientific › peer-review

11 Citations (Scopus)

Abstract

SARS-CoV-2 is evolving with increased transmission, host range, pathogenicity, and virulence. The original and mutant viruses escape host innate (Interferon) immunity and adaptive (Antibody) immunity, emphasizing unmet needs for high-yield, commercial-scale manufacturing to produce inexpensive vaccines/boosters for global/equitable distribution. We developed DYAI-100A85, a SARS-CoV-2 spike receptor binding domain (RBD) subunit antigen vaccine expressed in genetically modified thermophilic filamentous fungus, Thermothelomyces heterothallica C1, and secreted at high levels into fermentation medium. The RBD-C-tag antigen strongly binds ACE2 receptors in vitro. Alhydrogel^®‘85’-adjuvanted RDB-C-tag-based vaccine candidate (DYAI-100A85) demonstrates strong immunogenicity, and antiviral efficacy, including in vivo protection against lethal intranasal SARS-CoV-2 (D614G) challenge in human ACE2-transgenic mice. No loss of body weight or adverse events occurred. DYAI-100A85 also demonstrates excellent safety profile in repeat-dose GLP toxicity study. In summary, subcutaneous prime/boost DYAI-100A85 inoculation induces high titers of RBD-specific neutralizing antibodies and protection of hACE2-transgenic mice against lethal challenge with SARS-CoV-2. Given its demonstrated safety, efficacy, and low production cost, vaccine candidate DYAI-100 received regulatory approval to initiate a Phase 1 clinical trial to demonstrate its safety and efficacy in humans.

Original language	English
Article number	2119
Journal	Vaccines
Volume	10
Issue number	12
DOIs	https://doi.org/10.3390/vaccines10122119
Publication status	Published - 11 Dec 2022
MoE publication type	A1 Journal article-refereed

Funding

This work was funded in part by the Israel Institute for Biological Research, Ness-Ziona, Israel and by Dyadic, Jupiter International, Inc. Jupiter, FL, USA.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being
SDG 9 Industry, Innovation, and Infrastructure

Keywords

adjuvant
alum
aluminum-based vaccine adjuvants
antigen
baculovirus
BALB/c mice
biomanufacturing
C-tag
CHO-cell
COVID-19
dyadic
efficacy
glycans
glycoprotein
glycosylation
IgG
IgG1 and IgG2b
insect cells
intranasal challenge
K18-hACE-2 mice
Myceliophthora thermophila
neutralizing antibodies
pandemic
RBD
receptor binding domain
recombinant protein subunit vaccine
SARS-CoV-2
stability
Thermothelomyces heterothallica
toxicology
vaccine
variants of concern
virus
zoonotic

Access to Document

10.3390/vaccines10122119Licence: CC BY

Cite this

Nechooshtan, R., Ehrlich, S., Vitikainen, M., Makovitzki, A., Dor, E., Marcus, H., Hefetz, I., Pitel, S., Wiebe, M., Huuskonen, A., Cherry, L., Lupu, E., Sapir, Y., Holtzman, T., Aftalion, M., Gur, D., Tamir, H., Yahalom-Ronen, Y., Ramot, Y., ... Ophir, Y. (2022). Thermophilic Filamentous Fungus C1-Cell-Cloned SARS-CoV-2-Spike-RBD-Subunit-Vaccine Adjuvanted with Aldydrogel^®85 Protects K18-hACE2 Mice against Lethal Virus Challenge. Vaccines, 10(12), Article 2119. https://doi.org/10.3390/vaccines10122119

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title = "Thermophilic Filamentous Fungus C1-Cell-Cloned SARS-CoV-2-Spike-RBD-Subunit-Vaccine Adjuvanted with Aldydrogel{\textregistered}85 Protects K18-hACE2 Mice against Lethal Virus Challenge",

abstract = "SARS-CoV-2 is evolving with increased transmission, host range, pathogenicity, and virulence. The original and mutant viruses escape host innate (Interferon) immunity and adaptive (Antibody) immunity, emphasizing unmet needs for high-yield, commercial-scale manufacturing to produce inexpensive vaccines/boosters for global/equitable distribution. We developed DYAI-100A85, a SARS-CoV-2 spike receptor binding domain (RBD) subunit antigen vaccine expressed in genetically modified thermophilic filamentous fungus, Thermothelomyces heterothallica C1, and secreted at high levels into fermentation medium. The RBD-C-tag antigen strongly binds ACE2 receptors in vitro. Alhydrogel{\textregistered}{\textquoteleft}85{\textquoteright}-adjuvanted RDB-C-tag-based vaccine candidate (DYAI-100A85) demonstrates strong immunogenicity, and antiviral efficacy, including in vivo protection against lethal intranasal SARS-CoV-2 (D614G) challenge in human ACE2-transgenic mice. No loss of body weight or adverse events occurred. DYAI-100A85 also demonstrates excellent safety profile in repeat-dose GLP toxicity study. In summary, subcutaneous prime/boost DYAI-100A85 inoculation induces high titers of RBD-specific neutralizing antibodies and protection of hACE2-transgenic mice against lethal challenge with SARS-CoV-2. Given its demonstrated safety, efficacy, and low production cost, vaccine candidate DYAI-100 received regulatory approval to initiate a Phase 1 clinical trial to demonstrate its safety and efficacy in humans.",

keywords = "adjuvant, alum, aluminum-based vaccine adjuvants, antigen, baculovirus, BALB/c mice, biomanufacturing, C-tag, CHO-cell, COVID-19, dyadic, efficacy, glycans, glycoprotein, glycosylation, IgG, IgG1 and IgG2b, insect cells, intranasal challenge, K18-hACE-2 mice, Myceliophthora thermophila, neutralizing antibodies, pandemic, RBD, receptor binding domain, recombinant protein subunit vaccine, SARS-CoV-2, stability, Thermothelomyces heterothallica, toxicology, vaccine, variants of concern, virus, zoonotic",

author = "Ram Nechooshtan and Sharon Ehrlich and Marika Vitikainen and Arik Makovitzki and Eyal Dor and Hadar Marcus and Idan Hefetz and Shani Pitel and Marilyn Wiebe and Anne Huuskonen and Lilach Cherry and Edith Lupu and Yehuda Sapir and Tzvi Holtzman and Moshe Aftalion and David Gur and Hadas Tamir and Yfat Yahalom-Ronen and Yuval Ramot and Noam Kronfeld and David Zarling and Anne Vallerga and Ronen Tchelet and Abraham Nyska and Markku Saloheimo and Mark Emalfarb and Yakir Ophir",

note = "Publisher Copyright: {\textcopyright} 2022 by the authors.",

year = "2022",

month = dec,

day = "11",

doi = "10.3390/vaccines10122119",

language = "English",

volume = "10",

journal = "Vaccines",

issn = "2076-393X",

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Nechooshtan, R, Ehrlich, S, Vitikainen, M, Makovitzki, A, Dor, E, Marcus, H, Hefetz, I, Pitel, S, Wiebe, M , Huuskonen, A, Cherry, L, Lupu, E, Sapir, Y, Holtzman, T, Aftalion, M, Gur, D, Tamir, H, Yahalom-Ronen, Y, Ramot, Y, Kronfeld, N, Zarling, D, Vallerga, A, Tchelet, R, Nyska, A, Saloheimo, M, Emalfarb, M & Ophir, Y 2022, 'Thermophilic Filamentous Fungus C1-Cell-Cloned SARS-CoV-2-Spike-RBD-Subunit-Vaccine Adjuvanted with Aldydrogel^®85 Protects K18-hACE2 Mice against Lethal Virus Challenge', Vaccines, vol. 10, no. 12, 2119. https://doi.org/10.3390/vaccines10122119

TY - JOUR

T1 - Thermophilic Filamentous Fungus C1-Cell-Cloned SARS-CoV-2-Spike-RBD-Subunit-Vaccine Adjuvanted with Aldydrogel®85 Protects K18-hACE2 Mice against Lethal Virus Challenge

AU - Nechooshtan, Ram

AU - Ehrlich, Sharon

AU - Vitikainen, Marika

AU - Makovitzki, Arik

AU - Dor, Eyal

AU - Marcus, Hadar

AU - Hefetz, Idan

AU - Pitel, Shani

AU - Wiebe, Marilyn

AU - Huuskonen, Anne

AU - Cherry, Lilach

AU - Lupu, Edith

AU - Sapir, Yehuda

AU - Holtzman, Tzvi

AU - Aftalion, Moshe

AU - Gur, David

AU - Tamir, Hadas

AU - Yahalom-Ronen, Yfat

AU - Ramot, Yuval

AU - Kronfeld, Noam

AU - Zarling, David

AU - Vallerga, Anne

AU - Tchelet, Ronen

AU - Nyska, Abraham

AU - Saloheimo, Markku

AU - Emalfarb, Mark

AU - Ophir, Yakir

PY - 2022/12/11

Y1 - 2022/12/11

N2 - SARS-CoV-2 is evolving with increased transmission, host range, pathogenicity, and virulence. The original and mutant viruses escape host innate (Interferon) immunity and adaptive (Antibody) immunity, emphasizing unmet needs for high-yield, commercial-scale manufacturing to produce inexpensive vaccines/boosters for global/equitable distribution. We developed DYAI-100A85, a SARS-CoV-2 spike receptor binding domain (RBD) subunit antigen vaccine expressed in genetically modified thermophilic filamentous fungus, Thermothelomyces heterothallica C1, and secreted at high levels into fermentation medium. The RBD-C-tag antigen strongly binds ACE2 receptors in vitro. Alhydrogel®‘85’-adjuvanted RDB-C-tag-based vaccine candidate (DYAI-100A85) demonstrates strong immunogenicity, and antiviral efficacy, including in vivo protection against lethal intranasal SARS-CoV-2 (D614G) challenge in human ACE2-transgenic mice. No loss of body weight or adverse events occurred. DYAI-100A85 also demonstrates excellent safety profile in repeat-dose GLP toxicity study. In summary, subcutaneous prime/boost DYAI-100A85 inoculation induces high titers of RBD-specific neutralizing antibodies and protection of hACE2-transgenic mice against lethal challenge with SARS-CoV-2. Given its demonstrated safety, efficacy, and low production cost, vaccine candidate DYAI-100 received regulatory approval to initiate a Phase 1 clinical trial to demonstrate its safety and efficacy in humans.

AB - SARS-CoV-2 is evolving with increased transmission, host range, pathogenicity, and virulence. The original and mutant viruses escape host innate (Interferon) immunity and adaptive (Antibody) immunity, emphasizing unmet needs for high-yield, commercial-scale manufacturing to produce inexpensive vaccines/boosters for global/equitable distribution. We developed DYAI-100A85, a SARS-CoV-2 spike receptor binding domain (RBD) subunit antigen vaccine expressed in genetically modified thermophilic filamentous fungus, Thermothelomyces heterothallica C1, and secreted at high levels into fermentation medium. The RBD-C-tag antigen strongly binds ACE2 receptors in vitro. Alhydrogel®‘85’-adjuvanted RDB-C-tag-based vaccine candidate (DYAI-100A85) demonstrates strong immunogenicity, and antiviral efficacy, including in vivo protection against lethal intranasal SARS-CoV-2 (D614G) challenge in human ACE2-transgenic mice. No loss of body weight or adverse events occurred. DYAI-100A85 also demonstrates excellent safety profile in repeat-dose GLP toxicity study. In summary, subcutaneous prime/boost DYAI-100A85 inoculation induces high titers of RBD-specific neutralizing antibodies and protection of hACE2-transgenic mice against lethal challenge with SARS-CoV-2. Given its demonstrated safety, efficacy, and low production cost, vaccine candidate DYAI-100 received regulatory approval to initiate a Phase 1 clinical trial to demonstrate its safety and efficacy in humans.

KW - adjuvant

KW - alum

KW - aluminum-based vaccine adjuvants

KW - antigen

KW - baculovirus

KW - BALB/c mice

KW - biomanufacturing

KW - C-tag

KW - CHO-cell

KW - COVID-19

KW - dyadic

KW - efficacy

KW - glycans

KW - glycoprotein

KW - glycosylation

KW - IgG

KW - IgG1 and IgG2b

KW - insect cells

KW - intranasal challenge

KW - K18-hACE-2 mice

KW - Myceliophthora thermophila

KW - neutralizing antibodies

KW - pandemic

KW - RBD

KW - receptor binding domain

KW - recombinant protein subunit vaccine

KW - SARS-CoV-2

KW - stability

KW - Thermothelomyces heterothallica

KW - toxicology

KW - vaccine

KW - variants of concern

KW - virus

KW - zoonotic

UR - https://www.scopus.com/pages/publications/85144644532

U2 - 10.3390/vaccines10122119

DO - 10.3390/vaccines10122119

M3 - Article

C2 - 36560529

AN - SCOPUS:85144644532

SN - 2076-393X

VL - 10

JO - Vaccines

JF - Vaccines

IS - 12

M1 - 2119

ER -