A5014252803- Transgenic Plants and Applications
- Toxin Mechanisms and Immunotoxins
- Monoclonal and Polyclonal Antibodies Research
- SARS-CoV-2 and COVID-19 Research
- Viral Infectious Diseases and Gene Expression in Insects
- Plant tissue culture and regeneration
- Glycosylation and Glycoproteins Research
- Microbial Inactivation Methods
BOKU University
2021-2024
Monoclonal antibodies (mAbs) that efficiently neutralize SARS-CoV-2 have been developed at an unprecedented speed. Notwithstanding, there is a vague understanding of the various Ab functions induced beyond antigen binding by heavy-chain constant domain. To explore diverse roles Abs in immunity, we expressed spike protein (SP) mAb (H4) four IgG subclasses present human serum (IgG1-4) using glyco-engineered Nicotiana benthamiana plants. All subclasses, carrying identical antigen-binding site,...
Here, we expressed two neutralizing monoclonal antibodies (Abs) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; H4 and B38) in three formats: IgG1, IgA1 monomers (m), dimers (d) glycoengineered Nicotiana benthamiana plants. All six Ab variants assembled properly exhibited a largely homogeneous glycosylation profile. Despite modest variation antigen binding between formats, SARS-CoV-2 neutralization (NT) potency significantly increased the following manner: IgG1 <...
Immunoglobulin M (IgM) is the largest antibody isotype with unique features like extensive glycosylation and oligomerization. Major hurdles in characterizing its properties are difficulties production of well-defined multimers. Here we report expression two SARS-CoV-2 neutralizing monoclonal antibodies glycoengineered plants. Isotype switch from IgG1 to IgM resulted IgMs, composed 21 human protein subunits correctly assembled into pentamers. All four recombinant carried a highly reproducible...
This study describes a novel, neutralizing monoclonal antibody (mAb), 11D7, discovered by mouse immunization and hybridoma generation, against the parental Wuhan-Hu-1 RBD of SARS-CoV-2. We further developed this mAb into chimeric human IgG recombinantly expressed it in plants to produce with human-like, highly homogenous N-linked glycans that has potential impart greater potency safety as therapeutic. The epitope 11D7 was mapped competitive binding well-characterized mAbs, suggesting is...
Plants are being increasingly recognized for the production of complex human proteins, including monoclonal antibodies (mAbs). Various methods have been applied to boost recombinant expression, with DNA codon usage an important approach. Here, we transiently expressed three mAbs in Nicotiana benthamiana, namely one IgG3 and two IgM directed against SARS-CoV-2 as optimized(CO) non-codon optimized (NCO) variants. qRT-PCR exhibited significantly increased mRNA levels all CO variants compared...
Despite the unique advantages of IgG3 over other IgG subclasses, such as mediating enhanced effector functions and increased flexibility in antigen binding due to a long hinge region, therapeutic potential remains largely unexplored. This may be attributed difficulties recombinant expression reduced plasma half-life most allotypes. Here, we report plant two SARS-CoV-2 neutralizing monoclonal antibodies (mAbs) that exhibit high (P5C3) low (H4) binding. P5C3 H4-IgG1 mAbs were subclass-switched...
Plant-produced glycoproteins carry α1,3-linked core fucosylated N-glycans, and the reduction/elimination thereof often confers beneficial features (Strasser et al., 2008; Zeitlin 2011). Here, we evaluated possibility of altering fucosylation in planta by overexpressing a bacterial GDP-6-deoxy-d-lyxo-4-hexulose reductase (RMD), an enzyme negatively interfering with GDP-l-fucose biosynthesis pathway (von Horsten 2010). RMD gene from Pseudomonas aeruginosa (Figure S1) was transiently expressed...
Abstract Immunoglobulin M (IgM) is the largest antibody isotype with unique features like extensive glycosylation and oligomerization. Major hurdles in characterizing its properties are difficulties production of well-defined multimers. Here we report expression two SARS-CoV-2 neutralizing monoclonal antibodies glycoengineered plants. Isotype switch from IgG1 to IgM resulted pentameric IgMs, comprising correctly assembled 21 human protein subunits. All four recombinant carried a highly...