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abstract:
Cell engineering is the key technology of biopharmaceutical industry, and its application in pharmaceutical field has greatly developed biopharmaceutical industry, and cell engineering pharmacy has broad prospects. By sorting out and analyzing the relevant literature, this paper summarizes the related technologies in the field of cell engineering pharmacy and the significance and prospect of their application in biopharmaceutical industry.
Key words:
Cell engineering; Biopharmaceuticals; Animal cell engineering; Plant cell engineering; Transgenic; Reactor;
1, Overview of biopharmaceuticals and cell engineering
Biopharmaceutical is the comprehensive utilization of biotechnology. Effective components are separated from organisms, biological tissues, cells and body fluids to prepare products for prevention, treatment and diagnosis [1]. Natural biomaterials endow biopharmaceuticals with high safety, low side effects and high nutritional value. These remarkable advantages make biopharmaceuticals more and more popular, which is one of the important reasons for the continuous expansion of biopharmaceuticals market.
Cell engineering takes cells as the research object, uses the theory of cell and molecular biology to design and operate according to the needs, thus changing the genetic characteristics of cells, achieving the purpose of improving or creating new varieties, and finally extracting products beneficial to human beings after large-scale culture and reproduction. In industry, it mainly includes upstream engineering (including cell culture, gene manipulation and preservation) and downstream engineering (including the application of transformed cells in the production of biological products) [2]. Nowadays, cell engineering plays an irreplaceable role in biopharmaceutical industry.
2. Animal cell engineering pharmacy
2. 1. Overview and early development of animal cell engineering pharmacy
Animal cell engineering pharmacy can be traced back to 1950s, when animal cells were used to produce viruses, that is, animal cells were cultured in a bioreactor, and then attenuated or inactivated viruses were inoculated to produce vaccines [3]. The common technical process of animal cell culture is to first disperse animal tissues into single cells and cell groups (groups), then inoculate them into the culture medium for primary culture, and then subculture 10 ~ 50 generations to initially obtain the required cell lines. However, due to the low expression level of cells in nature, the products produced by this method are not only low in yield, but also high in cost. Therefore, early animal cell culture has not received enough attention.
2.2, hybridoma technology
The establishment of hybridoma technology in 1970s is a new milestone in the development of animal cell technology. With the application of hybridoma technology in the industrial field, various new products have appeared one after another, which is of great significance in the production of biological products for disease diagnosis and treatment [3]. From 65438 to 0984, the Nobel Prize in Physiology or Medicine was awarded to three scientists who founded the theory of antigen-selective antibody and invented monoclonal antibody technology. They proposed to fuse and screen B lymphocytes secreting specific antibodies with myeloma cells that can proliferate indefinitely to form hybridoma cells that can produce specific antibodies. The fused cells obtained by this method can stably produce monoclonal antibodies with strong specificity and high titer.
2.3, animal cell mass production technology
Large-scale production of animal cells refers to the cultivation of a large number of useful animal cells in a cell bioreactor under artificial conditions, which is the technology of producing drugs and the key technology of pharmaceutical industry. Because animal cells are highly sensitive to the changes of the external environment, the scale-up process of cell culture needs to be gradually scaled up from laboratory scale to production scale, and the difference of processes in each reactor has become a major technical challenge in the current scale-up process [4]. Using animal cells to produce biological products has become the main pillar of the global biological industry. At present, many biological agents obtained from animal cell culture are protein and antibodies.
2.4, animal bioreactor
Animal bioreactor can continuously obtain some protein needed by human beings from transgenic animals and produce protein industrially. According to different parts of protein, it can be divided into many types of bioreactors, such as blood bioreactor and salivary gland bioreactor. Scientists have found that mammary gland bioreactor has become the most promising development direction of bioreactor because the mammary gland of female animals can express recombinant protein efficiently and make some modifications. With the development of technology, the products of breast bioreactor have been extended to antithrombin, coagulation factor, human protein, lysozyme, superoxide dismutase, interferon and other enzymes or cytokines with high medical value. As a brand-new biological production mode, breast bioreactor will be widely used in biomedical industry because of its advantages of high yield and low cost in producing natural products [5].
2.5, animal nuclear transplantation
Animal nuclear transplantation also has a good prospect in cell engineering. The donor nucleus of an animal is taken out and injected into another enucleated oocyte in the metaphase of meiosis to change the genetic characteristics of the cell and produce new products, which are then cultured, propagated, purified and extracted in vitro, and finally used for disease treatment. The research on nuclear transfer of fish was the earliest in China. Tong Dizhou, the "father of China cloning", completed the world's first nuclear transfer of fish in 1960s. Later, Chinese scholars tried to carry out nuclear fusion experiments in other strains of fish, and used model animal zebrafish to reveal the molecular mechanism of reprogramming after nuclear transplantation, which achieved great research results and promoted the rapid development of nuclear transplantation technology and other related fields [6]. Nowadays, animal cell engineering is of great significance in the field of biopharmaceuticals. Because of the complexity of animal cell structure and the clarity of division of labor, animal cell engineering has great advantages.
3. Plant cell engineering pharmacy
3. 1. Overview and early development of plant cell engineering pharmacy
It is a traditional method to directly use plants as medicine or extract effective components from plants. With the maturity of technology, the process of treatment and extraction is becoming simpler and simpler. At present, many traditional Chinese medicines are produced in this way. However, this method is only suitable for plants that are easy to cultivate and reproduce quickly, and is not suitable for those plants with long growth cycle and difficult extraction, so it is subject to many restrictions. For example, Taxus chinensis containing anticancer components has suffered devastating damage because of large-scale logging [8].
Plant cell engineering pharmacy takes plant cells as the basic research unit, carries out a series of operations on plant cells, changes their biological characteristics, and finally achieves the purpose of improving or cultivating new varieties [9]. The application of plant cell and tissue culture has the advantages of less impurities, simple extraction, high content of effective components and short culture period. At present, plant cell engineering pharmacy is mainly embodied in tissue and cell culture, genetic characteristics transformation and transgenic plants.
3.2. Large-scale cultivation of plant cell engineering
In the 1950s, American scientists first proposed to use plants to extract natural medicines on a large scale. They obtained a large number of medicinal ingredients-furanone from multi-liter fermentor. As one of the countries with the longest history of plant medicine, the application of cell culture technology can help China traditional Chinese medicine play a greater role.
Salvia miltiorrhiza is a traditional Chinese medicine, which has the functions of promoting blood circulation, removing blood stasis, dredging menstruation and relieving pain. The main components-phenolic acids and diterpenoids-are mainly used to treat cardiovascular diseases. At present, due to the low content of effective components and slow growth of Salvia miltiorrhiza, the wild Salvia miltiorrhiza resources have been destroyed on a large scale, and the quality of varieties cultivated in different places is uneven, which is difficult to meet the market demand in quantity and quality [10]. The experimental study showed that the hairy roots of Salvia miltiorrhiza were cultured in a special plant tissue reactor with the scale of 10L, and the multiplication times of fresh weight were as high as 240 times in just 50 days, and the contents of various effective components were also greatly improved. This method is very suitable for the growth and product accumulation of hairy roots of Salvia miltiorrhiza, and avoids the pollution of pesticides and other substances.
3.3, plant transgenic technology
Compared with transgenic animals, transgenic plants have unique advantages: on the one hand, plant cells have totipotency, simple cell culture conditions and easy survival; On the other hand, foreign genes entering plants can accumulate beneficial genes in the process of hybridization with other plants for optimal expression. Transgenic plants can also be used to produce vaccines. Plants can be used as bioreactors, introducing vectors carrying antigen genes into recipient cells, and expressing and modifying such specific antigens in plants, thus becoming protein with immune activity. Bananas, carrots, potatoes, etc. Can be used as recipient plants. Some plant vaccines with transformation coding genes, such as HBsAg, LTB and Norwalk virus, have been used to prevent and treat hepatitis B and bacterial diarrhea. In biological and clinical trials, it showed a good immune response. Compared with the traditional vaccine, it has the advantages of low production cost, high success rate and easy to form large-scale production. Although the research of plant transgenic vaccine is still in its infancy, the biological experiment of transgenic plants reported in China has achieved certain results [1 1], which has become an important progress in China's pharmaceutical industry.
3.4, plant bioreactor
Plant bioreactor, also known as "plant gene pharmaceutical factory". This technology broadens the source of medicinal protein and vaccine, reduces the cost, expands the scale of biopharmaceutical industry, and has great commercial value. The research and development of plant bioreactor is of great significance to seize the commanding heights of bio-economy in the world. Many developed countries have included the research and development of plant bioreactors in the national strategic plan of key biotechnology research [12]. China began to develop nuclear power plants as reactors in 1990s. At present, there is still a certain gap between the research and investment of plant bioreactor and developed countries. Great progress has been made since the National Ninth Five-Year Plan gave policy support to this project [13].
4. Significance and prospect of cell engineering pharmacy.
It is of great significance to study the research progress and prospect of cell engineering pharmacy for the development of pharmaceutical industry. According to statistics, 50% of pharmaceutical products in the world come from cell engineering pharmacy, in which plant cell extracts and animal cell extracts each account for about 1/2. Cell engineering plays an important role in biopharmaceutical industry and provides a technical basis for new drug development. It is widely used to treat immune diseases, improve curative effect and drug innovation [8]. The research of cell engineering pharmacy is constantly making breakthroughs, and its influence and prospect are increasingly apparent. Nowadays, biopharmaceuticals and cell engineering have been closely linked. With the wide application of cell engineering technology in biopharmaceutical production, biopharmaceutical industry has developed rapidly and achieved great economic benefits [14].
With the emergence and update of more emerging technologies, we can make full use of various technical platforms to find the best research scheme in the future research and development of cell engineering drugs. The combination with other related fields will also better promote the development of biopharmaceuticals in China. In the past half century, cell engineering pharmacy has developed rapidly and achieved many research results in the medical field. Therefore, during the Tenth Five-Year Plan period, we should pay more attention to strategic emerging industries and further accelerate and expand the development of a new generation of biotechnology.
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