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Application of the iPLUS non-coding sequence in improving biopharmaceuticals production
Inês Reis-Claro1, Maria Inês Silva1, Ana Moutinho1
1Gene Regulation, i3S-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.
Frontiers in Bioengineering and Biotechnology
|February 21, 2024
Summary
A novel regulatory element, iPLUS, significantly boosts recombinant protein and mRNA production. This discovery enhances the efficiency of producing biological therapeutics, including monoclonal antibodies and mRNA vaccines, across various systems.
Area of Science:
- Biotechnology and Biopharmaceutical Production
- Molecular Biology and Gene Expression Regulation
Background:
- The biotechnology sector is rapidly expanding, with a focus on recombinant protein production for therapeutic applications.
- 3' untranslated region (3'UTR) cis-regulatory elements play a crucial role in controlling mRNA and protein expression levels.
Purpose of the Study:
- To investigate the function of 3'UTR cis-regulatory elements in enhancing mRNA and protein levels for biological therapeutics.
- To explore the regulatory potential of a universal sequence, iPLUS, for augmenting recombinant protein production.
Main Methods:
- Incorporation of the iPLUS sequence into expression vectors for monoclonal antibodies and reporter proteins.
- Utilized mammalian cell lines (ExpiCHO) and yeast Pichia pastoris for protein production experiments.
- Assessed the impact of iPLUS and its variant (iPLUSv2) on mRNA and protein levels in mRNA vaccine contexts (Spike, MAGEC2).
Main Results:
- iPLUS increased trastuzumab production by 2-fold in ExpiCHO cells.
- iPLUS (3x) and iPLUSv2 enhanced reporter protein production by over 100-fold in Pichia pastoris.
- iPLUSv2 doubled the production of Spike and MAGEC2 in mRNA vaccine applications.
Conclusions:
- The iPLUS sequence is a versatile tool for significantly increasing recombinant protein and mRNA levels across diverse biological systems.
- iPLUS demonstrates potential as a valuable asset for optimizing production processes in the biopharmaceutical industry, including mRNA vaccines.
- This study highlights the importance of non-coding sequences in gene expression control for biomanufacturing.
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