Recombinant protein production in plants

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Plants and plant cells are used to produce many different recombinant proteins, including vaccine subunits, antibodies and antibody fragments, and offer unique advantages in terms of production time, environmental impact, scalability and overall cost. Among the successfully produced target proteins were also some with high structural complexity, such as secretory IgAs, which pose a challenge for recombinant production in mammalian cells and often require additional technological solutions, such as reassociation in vitro. Overall, it thus appears that the plant cellular machinery is well suited for the expression and assembly of SIgAs and other complex multimeric proteins in functional form. However, the overall yield and in particular the proportion of fully assembled protein still varies depending on the protein. It is therefore desirable to further optimize plants accordingly and develop production lines that are ideally suited to synthesize complex proteins and lead to a higher yield of fully assembled and functional proteins. Technology: The endoplasmic reticulum (ER) is a central cell organelle for the synthesis of complex proteins, which are subsequently released into the lumen of the ER where they are folded with the help of chaperones and undergo some posttranslational modifications. The ER is well developed in most cells specialized for protein synthesis, and there is evidence in the literature that expansion of the ER increases ER resilience, supports a high protein production rate and alleviates ERstress. In the present invention, mutations were inserted in one to three N. benthamiana genes for CTP:phosphatidate cytidylyltransferase (CCT), so that a shortened version of the enzyme is produced, which is constantly present in an active conformation. CCT catalyzes a rate-limiting step for the synthesis of phosphatidylcholine (PC), which is an essential building block for ER membranes and causes the expansion of the ER. It has been shown that this expansion results in a higher yield of fully assembled and functional SIgA in the mutant plant lines, thus removing a major obstacle to realizing the full potential of SIgA for clinical use, both in humans and animals.

Use CasesContent extracted from patent full text and abstract with AI.

• Production of complex multimeric biologics (e.g., SIgA) in plants
• Cost-efficient R&D and vaccine/antibody prototyping platform

BenefitsContent extracted from patent full text and abstract with AI.

• Increased yield of functional (fully assembled) SlgA antibodies from plants
• Safe production of recombinant proteins on a large scale
• Optimized plant line for complex protein synthesis
• Cost-efficient manufacturing process

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Patent Abstract

The present invention refers to an engineered, non-naturally occurring nucleotide sequence encoding a constitutively active variant of a CTP-phosphocholine- cytidylyltransferase (CCT), wherein said nucleotide sequence encodes the catalytic domain of said CCT, comprises a frameshift mutation within 1 to 20 codons upstream of the nucleotide sequence encoding the lipid sensing domain of the CCT, or 1 to 15 codons downstream of the nucleotide sequence encoding the catalytic domain of the CCT, and optionally is under control of its endogenous promoter, and optionally encodes 1 to 15 amino acids of the lipid sensing domain of said CCT.