Biopharming: Transforming Plant System for Pharmaceutical Protein Production

Dr. Sonali Uppalwar, Yash Bhoir, Rohit Patil, Aniket Banchede, Jatin Gotarne,

doi:10.5281/zenodo.17651679

Review Paper | Open Access
Volume 03 | Issue 11 | Article Id IJPS/250311362

Biopharming: Transforming Plant System for Pharmaceutical Protein Production
Dr. Sonali Uppalwar* Yash Bhoir Rohit Patil Aniket Banchede Jatin Gotarne
Dept of Ideal Institute of Pharmacy, Wada, Palghar Mh421303.

Abstract

Biopharming is a novel biotechnological process that involves genetically altering plants to function as natural factories for the manufacture of medicinal proteins and essential compounds. Mammals, microorganisms (bacteria and yeast), insects, plants, and transgenic animals are all now producing recombinant proteins in cultured cell-based systems (reviewed by Demain and Vaishnav). Transgenic plants' effective eukaryotic protein synthesis, great scalability, comparatively low production costs, and no environmental impact make them an appealing system for the expression and synthesis of a wide range of proteins and biomolecules. This paper offers a thorough summary of the ground-breaking potential of biopharming, highlighting the intricate processes required to harness plants' natural ability to produce complex therapeutic proteins in a cost-effective and scalable way. The benefits and drawbacks of this innovative strategy are reviewed in this review, which also looks at the genetic engineering techniques used to turn plants into bioreactors. Additionally, it explains the regulatory structures that regulate biopharming and emphasizes the critical role that quality control protocols play in ensuring the effectiveness and safety of the pharmaceutical proteins made with this innovative technique. By examining recent advancements and applications, this study sheds light on the quickly evolving subject of biopharming and its promising future for the pharmaceutical industry.

Keywords

protein synthesis, great scalability, comparatively low production costs

Introduction

Biopharming is a relatively young area that naturally blends the ideas of biotechnology and agriculture. This section offers a strong starting point by shedding light on the important ramifications of biopharming in relation to current pharmaceutical research and development. The introduction provides a comprehensive overview of the basic concepts of genetic engineering and plant biotechnology, enabling plants to be reengineered to serve as versatile bioreactors for the large-scale manufacture of complex pharmaceutical proteins and medicinal chemicals. This paper examines the groundbreaking potential of biopharming, which uses the intricate biological processes of plants to get around the drawbacks of conventional pharmaceutical production techniques. Additionally, this introduction emphasizes the significance of biopharming in addressing the growing global demand for conveniently accessible and reasonably priced medications, emphasizing its vital role in fostering advancements in the accessibility and affordability of necessary medical treatments.[1], [2], [3], [4] This section facilitates a comprehensive understanding of the intricate relationships among genetic engineering, plant biology, and pharmaceutical production by illuminating the innovative methods and state-of-the-art strategies that underpin biopharming. It further highlights the significance of biopharmaceuticals' extensive research and development efforts to facilitate the synthesis of a variety of therapeutic proteins and advance biopharmaceuticals, which will ultimately increase the accessibility and efficacy of life-saving treatments for a wide range of illnesses.

Genetic Engineering Methods:

In the field of plant biopharming, a number of genetic engineering techniques are undoubtedly employed to facilitate the integration of certain genes into the plant genome and ensure the consistent expression of the required proteins. Modifying plant systems to serve as efficient bioreactors for the manufacture of therapeutic proteins requires the use of these techniques.[5] The following are some of the main genetic engineering methods utilized in plant biopharmaceutics:

1.Agrobacterium-Mediated Transformation: This technique uses the naturally occurring soil bacterium Agrobacterium tumefaciens as a vector to introduce desired genes into the plant genome. The Agrobacterium introduces the foreign gene into the plant cell, where it combines with the host genome to create the desired protein with consistent expression.[6]

Figure 1: Agrobacterium-Mediated Transformation.

Gene Gun or Particle Bombardment: This technique involves blasting tiny particles coated with the desired target gene into plant tissues. The high-velocity particles penetrate the cell walls and enable the foreign DNA to integrate with the plant genome, delivering it into the nucleus of the plant.[4], [7]

Figure 2: Gene Gun or Particle Bombardment.

Viral Vector-Mediated Gene Transfer: This technique entails creating specific viruses that carry and insert the targeted genes into plant cells. The viral vectors successfully transfer the foreign genetic material, enabling its incorporation into the genome of the host plant and the subsequent synthesis of the target protein.[8]

Figure 3: Viral Vector-Mediated Gene Transfer.

Protoplast Fusion and Microinjection: Protoplasts—plant cells with their cell walls removed—can be fused together to combine their genetic material. This facilitates the transfer of desirable genes between different plant species. Microinjection allows the precise integration and expression of the target genes by employing a tiny needle to inject foreign DNA straight into the nucleus of a plant cell.[9], [10]

Figure 4: Protoplast Fusion and Microinjection.

CRISPR/Cas9 Genome Editing: The CRISPR/Cas9 system allows for precision genome editing by focusing on specific DNA sequences within the plant genome. This technique affects metabolic pathways to increase the synthesis of medicinal substances or adds, removes, or modifies genes to facilitate the production of desired proteins.

Figure 5: CRISPR/Cas9 Genome Editing.

These genetic engineering techniques ensure consistent and efficient production of the desired proteins while facilitating the gradual integration of foreign genes into the plant genome. These cutting-edge techniques enable researchers to alter plant systems to produce valuable pharmaceutical proteins with enhanced yields and purity, hence increasing the potential uses of plant biopharming in the pharmaceutical industry.[11], [12], [13]

Criteria for selecting suitable plant species:

Selecting suitable plant species for plant biopharming and genetically modifying them to enhance their capacity to produce therapeutic proteins is an essential step in the biopharming process. Plant species selection, genetic alteration techniques, and regulatory considerations all affect the success of biopharming.[14], [15], [16] Below is a summary of the selection criteria for plant species and the genetic modification process:

Biomass and Growth Rate: A good plant species should ideally produce a large quantity of biomass and develop rapidly. Consequently, for each unit of area and time, more therapeutic proteins will be generated.

Genetic Amenable: The plant should be genetically receptive to transformation, which should make it easy for it to absorb and express foreign DNA. Genetic engineering can be applied more easily to some plants than others.

Expression Capacity: The chosen plant species should have the capacity to produce and store the required proteins. This capacity may differ among plant species due to their distinct metabolic processes and capacities for protein synthesis.

Ease of Scale-Up: The plant must be easy to grow on a big scale for commercial biopharming to succeed. Adaptability to various environmental conditions and ease of cultivation are important considerations.

Containment and Environmental Impact: It is essential to consider the potential for gene flow to wild or closely related plant species. Selecting plants that are unlikely to cross-pollinate with their wild counterparts helps ease concerns about the environment.

Safety and Regulation: Respect for legal requirements and safety precautions is essential. Whether or whether plants are subject to stricter regulations may affect the species selection.

Genetic Modification Process to Enhance Capacity:

The Genetic Modification Process involves the following critical steps:[2], [17], [18], [19]

Isolation of Target Genes: The first step is to analyse the genes that generate the therapeutic protein of interest. A variety of sources, including artificial DNA constructs, animals, and other plants, can provide these genes.

Vector Construction: The target genes are inserted into the vectors, which are circular DNA segments. These vectors might be suitable gene delivery vehicles, such as plasmids.

Transformation: Two techniques are employed to introduce the vectors containing the target genes into plant cells: particle bombardment and Agrobacterium-mediated transformation. Through this process, the foreign genes are integrated into the plant's DNA.

Selection and Regeneration: Foreign genes that have been successfully incorporated into plant cells are selected following transformation. After that, these cells are grown back into whole plants using tissue culture techniques.

Verification of Transgene Expression: The modified plants are assessed to ensure that the desired medicinal protein is being expressed. In this, molecular analysis—such as PCR and protein quantification—is widely employed.

Optimization: To increase the levels of protein expression, genetic engineering can be further improved. Modifying promoter sequences, regulatory regions, or other genetic components may be necessary to boost protein synthesis.

Field Trials and Commercial Cultivation: Once the safe and effective generation of the target proteins by the genetically modified plants has been proven, they can be grown on a larger scale for controlled field trials and eventually commercial production.

Figure 6: Genetic Modification Process.

Advantages and Disadvantages of Plant Biopharming:

The Advantages and Disadvantages of Plant Biopharming include: [14], [15], [20], [21]

Advantages	Disadvantages
1.Economical method of production.	1. Issues and complications with the laws pertaining to genetically modified organisms (GMOs).
2.Increased capacity for output.	2. Potential environmental risks, such as gene transfer to wild plant populations.
3.Decreased risk of contamination and the spread of pathogens.	3. Limited protein expression in specific plant species.
4. Protein synthesis that is efficient and rapid.	4. Formulation, purification, and downstream processing challenges.
5. Adaptability in protein engineering customisation.	5. Concerns about the use of genetically modified plants from an ethical standpoint.
6.Potential for producing medicinal proteins.	6. Little knowledge of the long-term effects on ecosystems and biodiversity.

Regulatory Frameworks:

Biosafety Regulations: Governments and international organizations have created biosafety regulations to guarantee the safe use of genetically modified organisms (GMOs) in biotechnology and agriculture, including plant biopharming. The primary objectives of these regulations are to ensure the containment of genetically modified plants and to assess any possible environmental risks to prevent their accidental introduction into the natural environment.[17]

Biotechnology Regulations: Several countries have adopted specific biotechnology laws that regulate the research, development, and commercialization of biotechnological goods, including those derived from plant biopharming. These regulations, which are intended to ensure the marketability and traceability of biopharmaceutical products, usually include stringent approval processes, risk assessment requirements, and labelling specifications.[22]

Good Manufacturing Practice (GMP) Guidelines: Regulatory agencies like the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) enforce GMP standards for the manufacturing, packaging, and labelling of pharmaceutical goods, including those produced through plant biopharming. In order to ensure the consistent quality, safety, and effectiveness of pharmaceutical proteins, these guidelines emphasize the need for standardized production processes and quality control systems.[22], [23], [24]

Quality Control Measures:

Identity and Purity Testing: Pharmaceutical proteins produced in plant-based bioreactors should undergo stringent identification and purity testing procedures to guarantee their authenticity and purity. These procedures could involve molecular tests, chromatographic methods, and spectroscopic analyses.[25], [26]

Potency and Efficacy Assessments: Conduct comprehensive potency and efficacy testing, such as in vitro and in vivo bioassays, to assess the biological activity and therapeutic efficacy of pharmaceutical proteins and ensure that they fulfil established standards and specifications.[27]

Stability and Shelf-life Studies: Examine stability and shelf-life under various storage conditions, including temperature, humidity, and light exposure, to ascertain the best handling and storage conditions for maintaining the integrity and stability of the pharmaceutical proteins over time.[28]

Contamination Control Measures: Adopt stringent contamination control protocols, such as routine environmental monitoring, aseptic handling techniques, and endotoxin testing, to prevent microbial contamination and the presence of endotoxins in the final pharmaceutical protein products.

Allergenicity Assessment: Conduct thorough evaluations of allergenicity using in silico predictive models, animal research, and human clinical trials to determine the potential allergy risks connected to therapeutic proteins derived from genetically modified plants. This measure helps ensure the safety of the finished product, particularly for those who are known to be allergic to or sensitive to specific proteins or compounds derived from plants.[29], [30]

Documentation and Record-keeping: Every stage of the production process, including cultivation, harvesting, downstream processing, and quality control testing, should be monitored and documented using comprehensive documentation and record-keeping systems to guarantee traceability and transparency in the manufacturing and distribution of pharmaceutical proteins.[31], [32]

Current Applications of Plant Biopharming in the Pharmaceutical Industry in various fields:

Vaccine Production: The creation of vaccines against a variety of infectious diseases, including hepatitis, influenza, and human papillomavirus (HPV), has demonstrated a notable use of the science of plant biopharmaceutics.[8], [33]

Monoclonal Antibody Production: The application of plant biopharming has enabled the manufacture of monoclonal antibodies that target specific diseases such as infectious diseases, autoimmune disorders, and cancer.[34], [35]

Enzyme Replacement Therapy: Lysosomal storage diseases and other inherited ailments may be treated with recombinant enzymes produced through plant biopharming. Less immunogenic and more bioavailable therapeutic enzymes than those used in conventional enzyme replacement treatments can be created with this technique.[12]

Orphan Drug Production: Utilizing plant biopharming has enabled the development of orphan pharmaceuticals, which target rare diseases and medical issues with few treatment options.[36], [37]

Antiviral Drug Development: Plant biopharmaceutics holds significant promise for the production of antiviral drugs, particularly for the treatment of emerging infectious diseases like COVID-19 and viral pandemics.[38], [39], [40]

Production of Diagnostic Reagents: Plant biopharming can help produce proteins and diagnostic reagents for use in medical testing and assays, allowing for accurate disease monitoring and diagnosis.[41]

Global Access to Biopharmaceuticals: Biopharmaceuticals may become more accessible globally if plants are grown in a range of geographic regions, particularly in developing nations without the infrastructure required for traditional pharmaceutical production.[9], [42], [43], [44].

CONCLUSION: As an organic and scalable platform for the synthesis of complex pharmacological proteins and biopharmaceuticals, plant biopharming is at the forefront of the groundbreaking integration of biotechnology and agriculture. Extensive research into state-of-the-art genetic engineering techniques, coupled with robust regulatory frameworks and stringent quality control procedures, underscores the potential of plant biopharming to revolutionize the pharmaceutical industry's protein synthesis methodology. With the current applications of plant biopharming in the development of vaccines, monoclonal antibodies, orphan medicines, and antiviral therapy, it is a major step toward resolving global health concerns and expanding access to necessary drugs globally.[1], [10], [12] Collaboration between researchers, biotechnologists, and regulatory agencies will be crucial to realizing the field's full potential as it advances and ensuring the sustainable and safe production of pharmaceutical proteins for improved healthcare around the world.[15], [29], [35].

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