ArticleFrontiers in plant science2016
Optimized Blanching Reduces the Host Cell Protein Content and Substantially Enhances the Recovery and Stability of Two Plant-Derived Malaria Vaccine Candidates.
Article in Frontiers in plant science, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 42 citations in OpenAlex.
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- Developing downstream processes for the purification of recombinant proteins and small molecules from Nicotiana benthamiana biomass.Plant biotechnology journal · 2026Review
- Design, optimization, production and activity testing of recombinant immunotoxins expressed in plants and plant cells for the treatment of monocytic leukemia.Bioengineered · 2023Article
- Risk assessment and bioburden evaluation of Agrobacterium tumefaciens-mediated transient protein expression in plants using the CaMV35S promoter.BMC biotechnology · 2023Article
- Product safety aspects of plant molecular farming.Frontiers in bioengineering and biotechnology · 2023Review
- The transient expression of recombinant proteins in plant cell packs facilitates stable isotope labelling for NMR spectroscopy.Plant biotechnology journal · 2022Article
- Enhanced HIV SOSIP Envelope yields in plants through transient co-expression of peptidyl-prolyl isomerase B and calreticulin chaperones and ER targeting.Scientific reports · 2022Article
- Optimising expression and extraction of recombinant proteins in plants.Frontiers in plant science · 2022Review
- Improving Recombinant Protein Recovery from Plant Tissue Using Heat Precipitation.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Contributions of the international plant science community to the fight against human infectious diseases - part 1: epidemic and pandemic diseases.Plant biotechnology journal · 2021Review
- Contributions of the international plant science community to the fight against infectious diseases in humans-part 2: Affordable drugs in edible plants for endemic and re-emerging diseases.Plant biotechnology journal · 2021Review
- Expression of Biofilm-Degrading Enzymes in Plants and Automated High-Throughput Activity Screening Using ExperimentalFrontiers in bioengineering and biotechnology · 2021Article
- Oral Immunization With a Plant HSP90-SAG1 Fusion Protein Produced in Tobacco Elicits Strong Immune Responses and Reduces Cyst Number and Clinical Signs of Toxoplasmosis in Mice.Frontiers in plant science · 2021Article
- Seasonal Weather Changes Affect the Yield and Quality of Recombinant Proteins Produced in Transgenic Tobacco Plants in a Greenhouse Setting.Frontiers in plant science · 2019Article
- A Combined Ultrafiltration/Diafiltration Step Facilitates the Purification of Cyanovirin-N From Transgenic Tobacco Extracts.Frontiers in bioengineering and biotechnology · 2018Article
- A Rapid Laser Probing Method Facilitates the Non-invasive and Contact-free Determination of Leaf Thermal Properties.Journal of visualized experiments : JoVE · 2017Article
- Comparison of Tobacco Host Cell Protein Removal Methods by Blanching Intact Plants or by Heat Treatment of Extracts.Journal of visualized experiments : JoVE · 2016Article
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Plants provide an advantageous expression platform for biopharmaceutical proteins because of their low pathogen burden and potential for inexpensive, large-scale production. However, the purification of target proteins can be challenging due to issues with extraction, the removal of host cell proteins (HCPs), and low expression levels. The heat treatment of crude extracts can reduce the quantity of HCPs by precipitation thus increasing the purity of the target protein and streamlining downstream purification. In the overall context of downstream process (DSP) development for plant-derived malaria vaccine candidates, we applied a design-of-experiments approach to enhance HCP precipitation from Nicotiana benthamiana extracts generated after transient expression, using temperatures in the 20-80°C range, pH values of 3.0-8.0 and incubation times of 0-60 min. We also investigated the recovery of two protein-based malaria vaccine candidates under these conditions and determined their stability in the heat-treated extract while it was maintained at room temperature for 24 h. The heat precipitation of HCPs was also carried out by blanching intact plants in water or buffer prior to extraction in a blender. Our data show that all the heat precipitation methods reduced the amount of HCP in the crude plant extracts by more than 80%, simplifying the subsequent DSP steps. Furthermore, when the heat treatment was performed at 80°C rather than 65°C, both malaria vaccine candidates were more stable after extraction and the recovery of both proteins increased by more than 30%.
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