TY  - JOUR
A1  - Knödler, Matthias
A1  - Rühl, Clemens
A1  - Emonts, Jessica
A1  - Buyel, Johannes Felix
T1  - Seasonal weather changes affect the yield and quality of recombinant proteins produced in transgenic tobacco plants in a greenhouse setting
T2  - Frontiers in Plant Science
N2  - Transgenic plants have the potential to produce recombinant proteins on an agricultural scale, with yields of several tons per year. The cost-effectiveness of transgenic plants increases if simple cultivation facilities such as greenhouses can be used for production. In such a setting, we expressed a novel affinity ligand based on the fluorescent protein DsRed, which we used as a carrier for the linear epitope ELDKWA from the HIV-neutralizing antibody 2F5. The DsRed-2F5-epitope (DFE) fusion protein was produced in 12 consecutive batches of transgenic tobacco (Nicotiana tabacum) plants over the course of 2 years and was purified using a combination of blanching and immobilized metal-ion affinity chromatography (IMAC). The average purity after IMAC was 57 ± 26% (n = 24) in terms of total soluble protein, but the average yield of pure DFE (12 mg kg−1) showed substantial variation (± 97 mg kg−1, n = 24) which correlated with seasonal changes. Specifically, we found that temperature peaks (>28°C) and intense illuminance (>45 klx h−1) were associated with lower DFE yields after purification, reflecting the loss of the epitope-containing C-terminus in up to 90% of the product. Whereas the weather factors were of limited use to predict product yields of individual harvests conducted for each batch (spaced by 1 week), the average batch yields were well approximated by simple linear regression models using two independent variables for prediction (illuminance and plant age). Interestingly, accumulation levels determined by fluorescence analysis were not affected by weather conditions but positively correlated with plant age, suggesting that the product was still expressed at high levels, but the extreme conditions affected its stability, albeit still preserving the fluorophore function. The efficient production of intact recombinant proteins in plants may therefore require adequate climate control and shading in greenhouses or even cultivation in fully controlled indoor farms.
KW  - batch reproducibility
KW  - environmental correlation
KW  - fluorescent protein carrier
KW  - greenhouse cultivation
KW  - plant molecular farming
Y1  - 2024
UR  - https://opus.bibliothek.fh-aachen.de/opus4/frontdoor/index/index/docId/11701
SN  - 1664-462X  (online-ressource)
IS  - 10
PB  - Frontiers Media
CY  - Lausanne
ER  -