Light-Induced Production of An Antibody Fragment and Malaria Vaccine Antigen from Chlamydomonas reinhardtii
Abstract
:1. Introduction
2. Experimental Section
2.1. Gene Constructs for αCD22 scFv and Pfs25
2.2. Cultivation of Recombinant Pfs25 and αCD22 scFv Chlamydomonas reinhardtii Strains
2.3. Effect of Light Duration and Light Intensity on Light-Induced Production of αCD22 scFv
2.4. Protein Extraction
2.5. Protein Analysis
2.6. FLAG Affinity Purification
2.7. Statistical Analysis
3. Results and Discussion
3.1. Algae Cultivation and Accumulation of αCD22scFv and Pfs25
3.2. Purification and Analysis of Recombinant Proteins
3.3. Light-Induced Accumulation of αCD22scFv
Photosynthetic Photon Flux (PPF) | Duration | 101 μmol m−2 s−1 | 300 μmol m−2 s−1 |
---|---|---|---|
Biomass (g) | 12 h | x 4.0 a ± 0.01 | x 4.9 a ± 1.5 |
24 h | x 4.0 a ± 0.01 | x 5.3 a ± 1.0 | |
TSP in algae extract (μg/mL) | 12 h | x 5939 a ± 257 | x 5485 a ± 802 |
24 h | x 6840 a ± 1167 | x 5248 a ± 415 | |
αCD22 scFv in wet biomass (μg/g) | 12 h | x 21.2 a ± 1.8 | x 26.6 a ± 4.3 |
24 h | x 35.1 b ± 5.8 | y 61.5 b ± 14 | |
αCD22 scFv volumetric conc. (μg/L) | 12 h | x 84.7 a ± 7.1 | x 135 a ± 66 |
24 h | x 140 a ± 20 | y 314 b ± 20 | |
αCD22 scFv (%TSP) | 12 h | x 0.07 a ± 0.01 | x 0.1 a ± 0.03 |
24 h | x 0.12 a ± 0.03 | y 0.23 b ± 0.05 |
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Munjal, N.; Garzon-Sanabria, A.J.; Quinones, K.W.; Gregory, J.; Nikolov, Z.L. Light-Induced Production of An Antibody Fragment and Malaria Vaccine Antigen from Chlamydomonas reinhardtii. Processes 2014, 2, 625-638. https://doi.org/10.3390/pr2030625
Munjal N, Garzon-Sanabria AJ, Quinones KW, Gregory J, Nikolov ZL. Light-Induced Production of An Antibody Fragment and Malaria Vaccine Antigen from Chlamydomonas reinhardtii. Processes. 2014; 2(3):625-638. https://doi.org/10.3390/pr2030625
Chicago/Turabian StyleMunjal, Neera, Andrea Juliana Garzon-Sanabria, Katelyn Wilson Quinones, James Gregory, and Zivko L. Nikolov. 2014. "Light-Induced Production of An Antibody Fragment and Malaria Vaccine Antigen from Chlamydomonas reinhardtii" Processes 2, no. 3: 625-638. https://doi.org/10.3390/pr2030625