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Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_5397_Библиотеки_им_академика_М_И_Перельмана

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5.12 Summary
Researchers have advanced microbial nanoparticle manufacturing by looking for particular properties in extremophiles and uncommon strains. For medication deliv­ery and medical uses, biocompatible nanoparticles with minimal cytotoxicity are created. A focus is on scaling up microbial nanoparticle synthesis for industrial usage, particularly multimetallic nanoparticles. Microbial synthesis contributes to environmental applications by eliminating heavy metals and contaminants in a sus­tainable manner. Tailoring characteristics, biomedical applications, nanoelectron­ics, and complex multimetallic architectures are all possibilities for microbial nanoparticle manufacturing in the future. Plant-mediated nanoparticle synthesis is a green approach that uses a variety of plant sources and has shown promise in medi­cation delivery, catalysis, and energy applications. Current research is geared at commercialization and industrial scale-up. The green technique of biopolymer and biomolecule-mediated nanoparticle production yields versatile nanoparticles with applications in medication administration, diagnostics, and environmental cleanup. Tailor-made nanoparticle creation, breakthroughs in medicinal applications, envi­ronmental remediation, and investigation of developing biopolymers for sustainable agriculture are all possibilities for the future. In conclusion, these approaches to sustainable nanoparticle synthesis have a wide range of applications, ranging from healthcare to environmental issues, with ongoing research focusing on large-scale production and commercialization.
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Chapter 6
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Production ofBiopharmaceuticals onGenetically Modied Organisms
SandipanJana, BarunDasGupta, AmitKar, AnirbitaGhosh, MoniharPervin, AsisBala, MotlalepulaG.Matsabisa, RajeswarDas, SomsubhraGhosh, PulokK.Mukherjee, andPallabK.Haldar
Abstract The production of biopharmaceuticals using genetically modied organ-
isms represents a transformative paradigm in the pharmaceutical industry. This abstract explores the advancements, challenges, and implications of employing genetically modied organisms for the synthesis of biopharmaceuticals. The genetic modication of organisms, ranging from bacteria and yeast to mammalian cells, enables the engineered expression of therapeutic proteins, antibodies, and vaccines. This review delves into the molecular techniques employed in genetically modied organisms to optimize protein production, the scalability of bioprocessing, and the regulatory landscape governing the use of genetically engineered platforms. Additionally, the chapter highlights the potential of genetically modied organisms to revolutionize the production of complex biologics and discusses the ethical and safety considerations associated with their widespread application in biopharma­ceutical manufacturing. The synthesis of biopharmaceuticals on genetically modi­ed organisms not only enhances production efciency but also contributes to the expanding landscape of precision medicine, heralding a new era in therapeutic inno­vation and healthcare delivery.
S. Jana (*) School of Pharmacy, The Neotia University, Sarisa, West Bengal, India
Department of Pharmaceutical Technology, Jadavpur University, Kolkata, West Bengal, India
B. D. Gupta School of Natural Product Studies, Jadavpur University, Kolkata, West Bengal, India
A. Kar · P. K. Mukherjee Institute of Bioresources and Sustainable Development, Autonomous Institute Under Department of Biotechnology, Imphal, Manipur, India e-mail: director.ibsd@nic.in
Ltd. 2024 S. Bose et al. (eds.), Concepts in Pharmaceutical Biotechnology and Drug Development, Interdisciplinary Biotechnological Advances,
https://doi.org/10.1007/978-981-97-1148-2_6
91© The Author(s), under exclusive license to Springer Nature Singapore Pte
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S. Jana et al.
Keywords Genetically modied organisms · Biopharmaceuticals · Upstream processing · Downstream processing
6.1 Biopharmaceuticals: AnIntroduction
In the past, the term “Biopharmaceuticals” referred to medications produced through biotechnological procedures utilizing molecular biology techniques. Today, these medications are widely used in various areas of medicine and are considered to be highly effective therapy options for numerous illnesses including cancer and metabolic disorders. Biopharmaceuticals encompass any benecial medication produced using genetically modied organisms, microbes, or com­pounds derived from biological processing (Kesik-Brodacka 2018). The produc- tion of these medications involves various biotechnology processes such as genetic engineering, recombinant human technology, gene transfer, and antibody produc­tion (Walsh 2000). The rst therapeutic protein and anti-diphtheria serum was introduced in 1894 by Farbwerke Hoechst to combat a catastrophic diphtheria epi­demic in Europe (Strohl and Knight 2009). Later developments included the pro­duction of heterologous insulin rened from pigs and cows and sold under the brand name “Iletin” and the cloning and expression of human insulin, somatosta­tin, and also growth hormone (HGH) in Escherichia coli in the late 1970s, which led to the creation of the rst recombinant human insulin (Bhatia and Goli 2018; Vecchio etal. 2018).
A. Ghosh · M. Pervin Department of Pharmaceutical Technology, Jadavpur University, Kolkata, West Bengal, India
A. Bala Division of Life Sciences, Institute of Advanced Study in Science and Technology (IASST), An Autonomous Institute Under Department of Science and Technology (Govt. of India) Vigyan Path, Guwahati, Assam, India e-mail: asisbala_ju@yhaoo.co.in
M. G. Matsabisa Department of Pharmacology, University of the Free State, Bloemfontein, South Africa e-mail: motlalepula.matsabisa@mrc.ac.za
R. Das · S. Ghosh School of Pharmacy, The Neotia University, Sarisa, West Bengal, India e-mail: rajeswar.das@tnu.in; somsubhra.ghosh@tnu.in
P. K. Haldar Department of Pharmaceutical Technology, Jadavpur University, Kolkata, West Bengal, India
School of Natural Product Studies, Jadavpur University, Kolkata, West Bengal, India
6 Production ofBiopharmaceuticals onGenetically Modied Organisms
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6.2 Production Processes
There are two stages of biopharmaceutical production—upstream processing and downstream processing.
6.2.1 Upstream Processing
Living cells such as those taken from patients that need to be genetically modied are used to produce biopharmaceutical drug substances using viral vectors. To cre­ate an ideal environment for cell development, it is important to choose the best cell line, culture medium, growth factors, and process optimization. Some crucial fac­tors to take into account are the regulation of temperature, pH, oxygen supply, and the sanitation of supplies and equipment utilized in the upkeep of an environment free of microorganisms. The procedure involves identifying and isolating the gene that produces the same protein expression in a healthy organism and selecting an appropriate host cell type for genetic engineering. Strict control and cGMP compli­ance over the handling of these living cells are critical to the outcome of the upstream production, which includes the amount of desired biopharmaceuticals produced, the type and quantity of biomolecular structural variants formed, and the kind and also the amount of specic impurities that need to be managed during purication (Jozala etal. 2016; Gronemeyer etal. 2014).
6.2.2 Downstream Processing
Downstream processing is a crucial part of the purication of biological products. It involves three steps: (1) removing most contaminants through purication, (2) elim­inating particular impurities and undesirable target biomolecule forms that might have developed during isolation and purication, and (3) polishing. Centrifugation, ltration, sedimentation, and otation are the primary techniques used to separate the products. However, continuous downstream processing has been known to increase process complexity rather than efciency in purication. Nevertheless, the benets of high output in continuous working units outweigh this drawback (Mehta 2019).
The current and most commonly used purication methods include continuous centrifugation, which involves retrieving the product from the extracellular media, cleaning and recovering it from inclusion bodies, clarifying cell homogenates, and harvesting precipitated protein. Filtration is another method used to eliminate byproducts and concentrate culture media containing the desired expressed biomol­ecules. Cell lysis by physical, chemical, and enzymatic methods is yet another
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method used for the production of biopharmaceuticals like recombinant proteins such as interferon-β, interleukin-2, protein C, and proinsulin, which are found to be produced as inclusion bodies inside the cells (Mehta 2019).
S. Jana et al.
6.3 Bacteria inBiopharmaceuticals Production
Bacteria can grow quickly on low-cost substrates, and researchers have extensively studied the method of producing recombinant proteins on a large scale in bioreac­tors. Microorganisms are a useful tool in producing recombinant proteins due to their metabolic power, diversity, and ability to adapt to various environmental con­ditions. The most commonly used microorganism is Escherichia coli, which has advanced genetics and a vast range of mutant host strains and cloning vectors. E. coli’s high growth rate, basic media requirement, ease of handling, high yield, and economic viability make it the most suitable for large-scale biopharmaceutical production, and E. coli can also produce recombinant proteins up to 80% of its dry matter (Sobolewska-Ruta and Zaleski 2019; Pham etal. 2019).
The incidence of diabetes is increasing globally, which is leading to an increase in demand for recombinant insulin therapies. For many years, insulin derived from cows and pigs was used to treat diabetes (Jozala etal. 2016). However, this was not the best option because many people experienced adverse reactions such as serum sickness. Genetic engineering made it possible to transfer genes across different biological spe­cies with ease and paved the way for efcient insulin manufacturing. Downstream processing of recombinant human insulin/analogue manufacture from E. coli inclu­sion bodies has been carried out. Recombinant human insulin derived from DNA was rst introduced by Eli Lilly & Company in 1982 and by Novo Nordisk in 1988 (Alyas etal. 2021). Humulin, the rst of several recombinant insulin to be licensed for use in general medicine, is made by Eli Lilly and Genentech. Recombinant insulin is still mainly produced in E. coli, where inclusion bodies are often used to express human insulin and its analogues. To generate sufcient amounts of proinsulin, recombinant E. coli is used. E. coli is transformed with proinsulin producing plasmids to create this recombinant protein. The plasmid includes proinsulin coding genes and a kanamycin mono sulphate resistance gene, which prolongs its survival rate (Siew and Zhang
2021). Next, the transformed cells are cultured on tryptic soy broth supplemented with
kanamycin mono sulphate, an antibiotic. E. coli cells that have not undergone trans­formation are eliminated by kanamycin mono sulphate. To maximize yields during product synthesis, the culture and fermentation conditions are closely monitored. Biologically active human insulin is produced by post-translational processing and thorough purication of the resultant medicinal product (Alyas etal. 2021).
The Pseudomonas genus is one of the most complex bacterial genera that researchers have investigated, showing a great deal of metabolic and physiological variety. A thorough investigation of Pseudomonas genomes and proteomics has led to a greater understanding of the system and molecular level of Pseudomonas syn­thetic processes. Due to its property of a vast array of genetic manipulation, fast growth rate, and the ability to secrete proteins, Pseudomonas uorescens has