Production of Pharmaceutical Grade Recombinant Native Aprotinin and Non-oxidized Aprotinin Variants Under Greenhouse and Field Conditions

Pogue, Gregory P.; Vojdani, Fakhrieh; Palmer, Kenneth E.; White, Earl L.; Haydon, Hugh; Bratcher, Barry

doi:10.1007/978-3-662-43836-7_5

Cited by 3 publications

(7 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Additional studies have demonstrated that GRFT can be stored in rice or in dried plant material for extended periods of time. This study provides evidence that it is possible to scale up current methods of viral-vector based GRFT manufacturing by expression in N. exclesiana, a cultivar that can be grown outdoors, unlike the conventional production host N. benthamiana (Pogue et al, 2010(Pogue et al, , 2014. We were able to express GRFT in N. excelsiana with an average of yield of 344 ± 129 mg/kg, which is approximately half of what is normally expressed in N. benthamiana plants.…”

Section: Discussionmentioning

confidence: 74%

“…The expression values and optimization related to the timing of infection were performed in a controlled indoor growth environment, which allows for evaluation relative to past N. benthamiana expression. N. excelsiana has the advantage of being developed for outdoor plant growth and should be able to be used in a more agricultural scale, allowing field production of GRFT (Pogue et al, 2010(Pogue et al, , 2014. In previous technoeconomic analysis GRFT production utilities accounted for over 34% of the total cost of goods (Alam et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

“…excelsiana is an interspecific hybrid derived from a cross between N. excelsior and N. benthamiana. Seed was kindly provided by Kentucky Bioprocessing (Pogue et al, 2010(Pogue et al, , 2014Shamloul et al, 2014).…”

Section: Plantsmentioning

confidence: 99%

See 2 more Smart Citations

In Preparation for Outdoor Pharming: Griffithsin Can Be Expressed in Nicotiana excelsiana and Retains Activity After Storage as Silage

Eapen

Cates

Mundell

et al. 2020

Front. Bioeng. Biotechnol.

Self Cite

View full text Add to dashboard Cite

Griffithsin is an algae-derived lectin with strong anti-viral activity against HIV and a positive safety profile. Multiple clinical studies are investigating griffithsin's utility as topical HIV microbicide. HIV microbicides are an extremely cost-sensitive market and plant-based griffithsin protein expression has the potential to meet those demands. The griffithsin product used in the clinic has been expressed and purified in N. benthamiana, using a TMV-based viral vector system, Geneware ® . Outdoor pharming of biopharmaceuticals would further alleviate startup costs for biotechnology firms and may allow broader product accessibility. Therefore, this study assessed expression in a hybrid tobacco line, N. excelsiana, that is susceptible to TMV-based viral vectors and can be grown outdoors. In addition to using this hybrid line we expand on methods for in planta storage of griffithsin in leafy plants by ensiling kilogram quantities of griffithsin. The ensiling process allows year-round biomanufacturing, minimal environmental-controlled storage, and reduces the industry need for multiple growth areas to maintain multi-product manufacturing of plant-based pharmaceuticals. This study shows that griffithsin can be expressed in N. excelsiana and is stable, recoverable, and active from ensiled tissue. These studies can pave the way for future plant-based pharmaceuticals to be expressed and stored in this manner.

show abstract

Section: Discussionmentioning

confidence: 74%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

In Preparation for Outdoor Pharming: Griffithsin Can Be Expressed in Nicotiana excelsiana and Retains Activity After Storage as Silage

Eapen

Cates

Mundell

et al. 2020

Front. Bioeng. Biotechnol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…The method combines the qualitative and quantitative assays of a protein mixture by polyacrylamide gel electrophoresis and simultaneous assessment of concentrations of the mixture components. It can also be used to develop special nutrition products containing pea seed proteins [24].…”

Section: Introductionmentioning

confidence: 99%

“…The target aprotinin and its impurities can be detected by gel electrophoresis as clearly as by chromatography [26]. This approach can also be proposed for monitoring product purification in various bioengineering processes (novel forms of food products [24] or novel sorbents for protein purification [27]) and in the development of pharmaceuticals [28].…”

Section: Introductionmentioning

confidence: 99%

On the Applicability of Electrophoresis for Protein Quantification

Dome

Akimenko

Бычков

et al. 2021

Preprint

View full text Add to dashboard Cite

Polyacrylamide gel electrophoresis (PAGE) is widely used for studying proteins and protein-containing objects. However, it is employed most frequently as a qualitative method rather than a quantitative one. In this paper, we show the feasibility of routine digital image acquisition and mathematical processing of electrophoregrams for protein quantification. Both the well-studied model protein molecules (bovine serum albumin) and more complex real-world protein-based products (casein-containing isolate for sports nutrition), which were subjected to mechanical activation in a planetary ball mill to obtain samples characterized by different protein denaturation degrees, were used as study objects. Protein quantification in the mechanically activated samples was carried out. The degree of destruction of individual protein was shown to be higher compared to that of protein-containing mixture after mechanical treatment for an identical amount of time. The methodological approach used in this study can serve as guidance for other researchers who would like to use electrophoresis for protein quantification both in individual form and in protein mixtures. The findings prove that photographic imaging of gels followed by mathematical data processing can be applied for analyzing the electrophoretic data.

show abstract

On the Applicability of Electrophoresis for Protein Quantification

et al. 2021

View full text Add to dashboard Cite

Polyacrylamide gel electrophoresis is widely used for studying proteins and protein-containing objects. However, it is employed most frequently as a qualitative method rather than a quantitative one. This paper shows the feasibility of routine digital image acquisition and mathematical processing of electropherograms for protein quantification when using vertical gel electrophoresis and Chrom & Spec software. Both the well-studied model protein molecules (bovine serum albumin) and more complex real-world protein-based products (casein-containing isolate for sports nutrition), which were subjected to mechanical activation in a planetary ball mill to obtain samples characterized by different protein denaturation degrees, were used as study objects. Protein quantification in the mechanically activated samples was carried out. The degree of destruction of individual protein was shown to be higher compared to that of the protein-containing mixture after mechanical treatment for an identical amount of time. The methodological approach used in this study can serve as guidance for other researchers who would like to use electrophoresis for protein quantification both in individual form and in protein mixtures. The findings prove that photographic imaging of gels followed by mathematical data processing can be applied for analyzing the electrophoretic data as an affordable, convenient and quick tool.

show abstract

Production of Pharmaceutical Grade Recombinant Native Aprotinin and Non-oxidized Aprotinin Variants Under Greenhouse and Field Conditions

Cited by 3 publications

References 34 publications

In Preparation for Outdoor Pharming: Griffithsin Can Be Expressed in Nicotiana excelsiana and Retains Activity After Storage as Silage

In Preparation for Outdoor Pharming: Griffithsin Can Be Expressed in Nicotiana excelsiana and Retains Activity After Storage as Silage

On the Applicability of Electrophoresis for Protein Quantification

On the Applicability of Electrophoresis for Protein Quantification

Contact Info

Product

Resources

About