Production of Recombinant Proteins from Transgenic Crops

Nikolov, Z̆ivko L.; Hammes, Daniel

doi:10.1007/978-94-017-2693-1_8

Cited by 20 publications

(7 citation statements)

References 12 publications

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“…Tobacco leaf is often used for agroinfiltration (Dillen et al, 1997;Kapila et al, 1997;Stoger et al, 2002;Vaquero et al, 1999Vaquero et al, , 2002Yang et al, 2000). However, tobacco leaves contain substances causing difficulty for purification, including alkaloids (Fischer et al, 2004) and phenolics (Nikolov and Hammes, 2002). Alternatively, lettuce was found to readily express the recombinant proteins b-glucuronidase (GUS) and green fluorescent protein (GFP) following agroinfiltration (Wroblewski et al, 2005) and lettuce has been stably transformed to produce an edible vaccine (Kapusta et al, 1999).…”

Section: Introductionmentioning

confidence: 98%

High‐level transient expression of recombinant protein in lettuce

Joh

Wróblewski

Ewing

et al. 2005

Biotech & Bioengineering

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Transient expression following agroinfiltration of plant tissue was investigated as a system for producing recombinant protein. As a model system, Agrobacterium tumefaciens containing the beta-glucuronidase (GUS) gene was vacuum infiltrated into lettuce leaf disks. Infiltration with a suspension of 10(9) colony forming units/mL followed by incubation for 72 h at 22 degrees C in continuous darkness produced a maximum of 0.16% GUS protein based on dry tissue or 1.1% GUS protein based on total soluble protein. This compares favorably to expression levels for commercially manufactured GUS protein from transgenic corn seeds. A. tumefaciens culture medium pH between 5.6 and 7.0 and surfactant concentrations < or = 100 ppm in the vacuum infiltration did not affect GUS expression, while infiltration with an A. tumefaciens density of 10(7) and 10(8) colony forming units/mL, incubation at 29 degrees C, and a surfactant concentration of 1,000 ppm significantly reduced expression. Incubation in continuous light caused lettuce to produce GUS protein more rapidly, but final levels did not exceed the GUS production in leaves incubated in continuous darkness after 72 h at 22 degrees C. The kinetics of GUS expression during incubation in continuous light and dark were represented well using a logistic model, with rate constants of 0.30 and 0.29/h, respectively. To semi-quantitatively measure the GUS expression in large numbers of leaf disks, a photometric enhancement of the standard histochemical staining method was developed. A linear relationship with an R2 value of 0.90 was determined between log10 (% leaf darkness) versus log10 (GUS activity). Although variability in expression level was observed, agroinfiltration appears to be a promising technology that could potentially be scaled up to produce high-value recombinant proteins in planta.

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Section: Introductionmentioning

confidence: 98%

High‐level transient expression of recombinant protein in lettuce

Joh

Wróblewski

Ewing

et al. 2005

Biotech & Bioengineering

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“…With respect to raw material cost, transgenic rice is expected to incur additional cost to cover expenses related regulatory compliance. 44–46 If the cost of transgenic rice is assumed to be 5 times greater than the production cost of commodity rice, the raw material cost of $1/kg would then account for 13% of the total manufacturing cost for the simulated process. For the egg‐white process, the raw material cost of $0.33/kg egg‐white would amount to 6% of the total cost.…”

Section: Resultsmentioning

confidence: 99%

Process evaluations and economic analyses of recombinant human lysozyme and hen egg‐white lysozyme purifications

Wilken

Nikolov

2011

Biotechnology Progress

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Human lysozyme and hen egg-white lysozyme have antibacterial, antiviral, and antifungal properties with numerous potential commercial applications. Currently, hen egg-white lysozyme dominates low cost applications but the recent high-level expression of human lysozyme in rice could provide an economical source of lysozyme. This work compares human lysozyme and hen egg-white lysozyme adsorption to the cation exchange resin, SP-Sepharose FF, and the effect of rice extract components on lysozyme purification. With one exception, the dynamic binding capacities of human lysozyme were lower than those of hen egg-white at pH 4.5, 6, and 7.5 with ionic strengths ranging from 0 to 100 mM (5-20 mS). Ionic strength and pH had a similar effect on the adsorption capacities, but human lysozyme was more sensitive to these two factors than hen egg-white lysozyme. In the presence of rice extract, the dynamic binding capacities of human and hen egg-white lysozymes were reduced by 20-30% and by 32-39% at pH 6. Hen egg-white lysozyme was used as a benchmark to compare the effectiveness of human lysozyme purification from transgenic rice extract. Process simulation and cost analyses for human lysozyme purification from rice and hen egg-white lysozyme purification from egg-white resulted in similar unit production costs at 1 ton per year scale.

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“…Antifungal proteins, such as chitinases and beta-1,3glucanases, have been genetically engineered to attempt expression in the maize kernels with the aim to prevent the growth of Aspergillus flavus and the production of aflatoxins (Duncan et al,1985;Wu et al, 1994;Wan et al, 1995). Transgenic maize will serve as bioreactors for producing various biomolecules with applications in food, feed and the pharmaceutical industry (Nikolov, 1999).…”

Section: Appendix I Maize Biotechnologymentioning

confidence: 99%

Section 2 - Soybean (GLYCINE MAX (L.) MARR.)

2006

Harmonisation of Regulatory Oversight in Biotechnology

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The OECD is a unique forum where the governments of 30 democracies work together to address the economic, social and environmental challenges of globalisation. The OECD is also at the forefront of efforts to understand and to help governments respond to new developments and concerns, such as corporate governance, the information economy and the challenges of an ageing population. The Organisation provides a setting where governments can compare policy experiences, seek answers to common problems, identify good practice and work to co-ordinate domestic and international policies.

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Production of Recombinant Proteins from Transgenic Crops

Cited by 20 publications

References 12 publications

High‐level transient expression of recombinant protein in lettuce

High‐level transient expression of recombinant protein in lettuce

Process evaluations and economic analyses of recombinant human lysozyme and hen egg‐white lysozyme purifications

Section 2 - Soybean (GLYCINE MAX (L.) MARR.)

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