Journal articles: 'Biopharmaceutical proteins'

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Liu, Shulei, and Benjamin L. Schulz. "Biopharmaceutical quality control with mass spectrometry." Bioanalysis 13, no. 16 (August 2021): 1275–91. http://dx.doi.org/10.4155/bio-2021-0123.

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Mass spectrometry (MS) is a powerful technique for protein identification, quantification and characterization that is widely applied in biochemical studies, and which can provide data on the quantity, structural integrity and post-translational modifications of proteins. It is therefore a versatile and widely used analytic tool for quality control of biopharmaceuticals, especially in quantifying host-cell protein impurities, identifying post-translation modifications and structural characterization of biopharmaceutical proteins. Here, we summarize recent advances in MS-based analyses of these key quality attributes of the biopharmaceutical development and manufacturing processes.

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Todorovic, Zoran, and Dragana Protic. "Bioethical issues in the development of biopharmaceuticals." Filozofija i drustvo 23, no. 4 (2012): 49–56. http://dx.doi.org/10.2298/fid1204049t.

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Development of biopharmaceuticals is a challenging issue in bioethics. Unlike conventional, small molecular weight drugs, biopharmaceuticals are proteins derived from DNA technology and hybrid techniques with complex three dimensional structures. Immunogenicity of biopharmaceuticals should always be tested in clinical settings due to low predictive value of preclinical animal models. However, non-human primates (NHP) and transgenic mice could be used to address certain aspects of immunogenicity. Substantial efforts have been made to reduce NHP use in biopharmaceutical drug development, e.g. study design improvements and changes in regulatory policy. In addition, several expert groups are active in this field (e.g. NC3Rs, BioSafe, and Biopharmaceutical Technical Group). Despite that, there is an increasing trend of use of NHP in preclinical safety testing of biopharmaceuticals, especially regarding monoclonal antibodies. Other potential bioethical issues related biopharmaceutical drug development are their cost/effectiveness ratio, clinical safety assessment, production of biosimilars, and comparison of their efficacy with placebo in countries without intention to market. Identification of the human genome has opened many new bioethical issues. Development of biopharmaceuticals is an important bioethical issue for several reasons. It connects all aspects of contemporary bioethics: bio?medicine (e.g. clinical trials in vulnerable subjects), animal welfare and the most recent ad?vances in biotechnology. In particular, biopharmaceutical drug development is a challenging issue regarding treatment of rare diseases.

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Barolo, Lorenzo, Raffaela M. Abbriano, Audrey S. Commault, Jestin George, Tim Kahlke, Michele Fabris, Matthew P. Padula, Angelo Lopez, Peter J. Ralph, and Mathieu Pernice. "Perspectives for Glyco-Engineering of Recombinant Biopharmaceuticals from Microalgae." Cells 9, no. 3 (March 5, 2020): 633. http://dx.doi.org/10.3390/cells9030633.

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Microalgae exhibit great potential for recombinant therapeutic protein production, due to lower production costs, immunity to human pathogens, and advanced genetic toolkits. However, a fundamental aspect to consider for recombinant biopharmaceutical production is the presence of correct post-translational modifications. Multiple recent studies focusing on glycosylation in microalgae have revealed unique species-specific patterns absent in humans. Glycosylation is particularly important for protein function and is directly responsible for recombinant biopharmaceutical immunogenicity. Therefore, it is necessary to fully characterise this key feature in microalgae before these organisms can be established as industrially relevant microbial biofactories. Here, we review the work done to date on production of recombinant biopharmaceuticals in microalgae, experimental and computational evidence for N- and O-glycosylation in diverse microalgal groups, established approaches for glyco-engineering, and perspectives for their application in microalgal systems. The insights from this review may be applied to future glyco-engineering attempts to humanize recombinant therapeutic proteins and to potentially obtain cheaper, fully functional biopharmaceuticals from microalgae.

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Zhang, Fangrong, Gesa Richter, Benjamin Bourgeois, Emil Spreitzer, Armin Moser, Andreas Keilbach, Petra Kotnik, and Tobias Madl. "A General Small-Angle X-ray Scattering-Based Screening Protocol for Studying Physical Stability of Protein Formulations." Pharmaceutics 14, no. 1 (December 28, 2021): 69. http://dx.doi.org/10.3390/pharmaceutics14010069.

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A fundamental step in developing a protein drug is the selection of a stable storage formulation that ensures efficacy of the drug and inhibits physiochemical degradation or aggregation. Here, we designed and evaluated a general workflow for screening of protein formulations based on small-angle X-ray scattering (SAXS). Our SAXS pipeline combines automated sample handling, temperature control, and fast data analysis and provides protein particle interaction information. SAXS, together with different methods including turbidity analysis, dynamic light scattering (DLS), and SDS-PAGE measurements, were used to obtain different parameters to provide high throughput screenings. Using a set of model proteins and biopharmaceuticals, we show that SAXS is complementary to dynamic light scattering (DLS), which is widely used in biopharmaceutical research and industry. We found that, compared to DLS, SAXS can provide a more sensitive measure for protein particle interactions, such as protein aggregation and repulsion. Moreover, we show that SAXS is compatible with a broader range of buffers, excipients, and protein concentrations and that in situ SAXS provides a sensitive measure for long-term protein stability. This workflow can enable future high-throughput analysis of proteins and biopharmaceuticals and can be integrated with well-established complementary physicochemical analysis pipelines in (biopharmaceutical) research and industry.

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Nielsen, Jens. "Production of biopharmaceutical proteins by yeast." Bioengineered 4, no. 4 (July 2013): 207–11. http://dx.doi.org/10.4161/bioe.22856.

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Webber, Matthew J., Eric A. Appel, Brittany Vinciguerra, Abel B. Cortinas, Lavanya S. Thapa, Siddharth Jhunjhunwala, Lyle Isaacs, Robert Langer, and Daniel G. Anderson. "Supramolecular PEGylation of biopharmaceuticals." Proceedings of the National Academy of Sciences 113, no. 50 (November 28, 2016): 14189–94. http://dx.doi.org/10.1073/pnas.1616639113.

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The covalent modification of therapeutic biomolecules has been broadly explored, leading to a number of clinically approved modified protein drugs. These modifications are typically intended to address challenges arising in biopharmaceutical practice by promoting improved stability and shelf life of therapeutic proteins in formulation, or modifying pharmacokinetics in the body. Toward these objectives, covalent modification with poly(ethylene glycol) (PEG) has been a common direction. Here, a platform approach to biopharmaceutical modification is described that relies on noncovalent, supramolecular host–guest interactions to endow proteins with prosthetic functionality. Specifically, a series of cucurbit[7]uril (CB[7])–PEG conjugates are shown to substantially increase the stability of three distinct protein drugs in formulation. Leveraging the known and high-affinity interaction between CB[7] and an N-terminal aromatic residue on one specific protein drug, insulin, further results in altering of its pharmacological properties in vivo by extending activity in a manner dependent on molecular weight of the attached PEG chain. Supramolecular modification of therapeutic proteins affords a noncovalent route to modify its properties, improving protein stability and activity as a formulation excipient. Furthermore, this offers a modular approach to append functionality to biopharmaceuticals by noncovalent modification with other molecules or polymers, for applications in formulation or therapy.

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Owczarek, B., A. Gerszberg, and K. Hnatuszko-Konka. "A Brief Reminder of Systems of Production and Chromatography-Based Recovery of Recombinant Protein Biopharmaceuticals." BioMed Research International 2019 (January 8, 2019): 1–13. http://dx.doi.org/10.1155/2019/4216060.

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Recombinant proteins are produced for various applications in laboratory and industrial settings. Among them, therapeutic applications have evolved into a mature field in recent years, affecting the face of contemporary medical treatment. This, in turn, has stimulated an ever-greater need for innovative technologies for the description, expression, and purification of recombinant protein biopharmaceuticals. Therefore, many biopharmaceuticals are synthesized in heterologous systems to obtain satisfactory yields that cannot be provided by natural sources. As more than 35 years has passed since the first recombinant biopharmaceutical (human insulin) successfully completed clinical trials in humans, we provide a brief review of the available prokaryotic and eukaryotic expression systems, listing the advantages and disadvantages of their use. Some examples of therapeutic proteins expressed in heterologous hosts are also provided. Moreover, technologies for the universal extraction of protein molecules are mentioned here, as is the methodology of their purification.

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Buyel, Johannes Felix, and Rainer Fischer. "Downstream processing of biopharmaceutical proteins produced in plants." Bioengineered 5, no. 2 (February 3, 2014): 138–42. http://dx.doi.org/10.4161/bioe.28061.

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Howard, John A. "Commercialization of Biopharmaceutical and Bioindustrial Proteins from Plants." Crop Science 45, no. 2 (March 2005): 468–72. http://dx.doi.org/10.2135/cropsci2005.0468.

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Creamer, Jessica S., Nathan J. Oborny, and Susan M. Lunte. "Recent advances in the analysis of therapeutic proteins by capillary and microchip electrophoresis." Anal. Methods 6, no. 15 (2014): 5427–49. http://dx.doi.org/10.1039/c4ay00447g.

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Hamrang, Zahra, Nicholas J. W. Rattray, and Alain Pluen. "Proteins behaving badly: emerging technologies in profiling biopharmaceutical aggregation." Trends in Biotechnology 31, no. 8 (August 2013): 448–58. http://dx.doi.org/10.1016/j.tibtech.2013.05.004.

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Tolbert, William R. "Manufacture of biopharmaceutical proteins by mammalian cell culture systems." Biotechnology Advances 8, no. 4 (January 1990): 729–39. http://dx.doi.org/10.1016/0734-9750(90)91994-r.

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Martin, Pauline L., and Gerhard F. Weinbauer. "Developmental Toxicity Testing of Biopharmaceuticals in Nonhuman Primates." International Journal of Toxicology 29, no. 6 (October 6, 2010): 552–68. http://dx.doi.org/10.1177/1091581810378896.

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Developmental toxicity studies for pharmaceutical safety testing are designed to evaluate potential adverse effects of drug treatment on pregnancy and on the developing embryo/fetus. Biopharmaceuticals present specific challenges for developmental toxicity testing because the pharmacology of these molecules, which are frequently human-specific proteins, is often restricted to humans and nonhuman primates (NHPs). For those species-restricted molecules, the only option for the evaluation of potential effects on development of the human biopharmaceutical is to use NHPs. This article reviews each of the stages of development in cynomolgus macaques (the most frequently used NHP) and the potential exposure of the embryo, fetus, and infant following administration of a biopharmaceutical during pregnancy and lactation. Because the purpose of the NHP developmental studies is to identify potential human risks, a comparison between macaque and human development and potential exposure has been made when possible. Understanding the potential exposure of the conceptus relative to critical periods in development is essential to designing a scientifically based study that adequately addresses human risks. Some options for NHP study designs, including the option of combining end points into a single study, and the pros and cons of each of the study options have been reviewed. Developmental studies for biopharmaceuticals in NHPs need to be optimally designed on a case-by-case basis taking into consideration the pharmacology of the molecule, the type of molecule (antibody or non-antibody), the potential exposure relative to the development of potential target organs, the clinical use, and the ethical considerations associated with the use of NHPs.

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Sasaki, Tetsuji, and Akiyoshi Taniguchi. "Development of a Non-protein and Lipid Medium Adopted Cell Line for Biopharmaceutical Recombinant Protein Expression." Open Biotechnology Journal 7, no. 1 (February 22, 2013): 1–6. http://dx.doi.org/10.2174/1874070701307010001.

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Recently, many biopharmaceuticals have been developed such as cytokines, growth factors, and antibodies. These recombinant proteins are mostly expressed by CHO cells. However, the culture medium of CHO cells requires the addition of serum, which can contain unknown biological substances such as viruses, or requires the addition of expensive growth factors. To avoid the risks of biological ingredients and to decrease the cost of biopharmaceutical production, we developed a non-protein and lipid medium adopted (NPLAd) CHO cell line using the adapted culture method. Our results indicated that autocrine EGF production and insulin addition are essential for NPLAd CHO cell growth. However, the rate of cell proliferation of NPLAd CHO cells was decreased compared with original CHO-K1 cells. The proliferation of NPLAd CHO cells was improved by GM3 addition, suggesting increased signaling efficiency of autocrine factors. No difference was found in the growth rate between original CHO-K1 and NPLAd CHO cells supplemented with insulin and GM3. The productivity of recombinant protein in NPLAd CHO cells was verified using secreting luciferase reporter system. As a result, luciferase activity in NPLAd CHO cells showed more than three times higher than in the original CHOK1 cells. The results suggested that this cell line could be useful for biopharmaceutical recombinant protein.

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Onoue, Satomi, Hiroki Suzuki, and Yoshiki Seto. "Formulation Approaches to Overcome Biopharmaceutical Limitations of Inhaled Peptides/Proteins." Current Pharmaceutical Design 21, no. 27 (September 17, 2015): 3867–74. http://dx.doi.org/10.2174/1381612821666150820110826.

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Rodger, Alison, and Doug Marshall. "Beginners guide to circular dichroism." Biochemist 43, no. 2 (March 26, 2021): 58–64. http://dx.doi.org/10.1042/bio_2020_105.

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Circular dichroism (CD) is used to give information about the chirality or handedness of molecular systems. It is particularly widely applied to determine the secondary structure of proteins such as biopharmaceutical products.

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Handl, Alina, Ángela I. López-Lorente, René Handrick, Boris Mizaikoff, and Friedemann Hesse. "Infrared attenuated total reflection and 2D fluorescence spectroscopy for the discrimination of differently aggregated monoclonal antibodies." Analyst 144, no. 21 (2019): 6334–41. http://dx.doi.org/10.1039/c9an00424f.

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Antibody aggregates may occur as undesirable by-products during manufacturing of biopharmaceutical proteins since parameters such as pH, temperature, ionic strength, protein concentration, oxygen, and shear forces can lead to aggregate formation.

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Bolje, Aljoša, and Stanislav Gobec. "Analytical Techniques for Structural Characterization of Proteins in Solid Pharmaceutical Forms: An Overview." Pharmaceutics 13, no. 4 (April 11, 2021): 534. http://dx.doi.org/10.3390/pharmaceutics13040534.

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Therapeutic proteins as biopharmaceuticals have emerged as a very important class of drugs for the treatment of many diseases. However, they are less stable compared to conventional pharmaceuticals. Their long-term stability in solid forms, which is critical for product performance, depends heavily on the retention of the native protein structure during the lyophilization (freeze-drying) process and, thereafter, in the solid state. Indeed, the biological function of proteins is directly related to the tertiary and secondary structure. Besides physical stability and biological activity, conformational stability (three-dimensional structure) is another important aspect when dealing with protein pharmaceuticals. Moreover, denaturation as loss of higher order structure is often a precursor to aggregation or chemical instability. Careful study of the physical and chemical properties of proteins in the dried state is therefore critical during biopharmaceutical drug development to deliver a final drug product with built-in quality that is safe, high-quality, efficient, and affordable for patients. This review provides an overview of common analytical techniques suitable for characterizing pharmaceutical protein powders, providing structural, and conformational information, as well as insights into dynamics. Such information can be very useful in formulation development, where selecting the best formulation for the drug can be quite a challenge.

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Castro, Leonor S., Guilherme S. Lobo, Patrícia Pereira, Mara G. Freire, Márcia C. Neves, and Augusto Q. Pedro. "Interferon-Based Biopharmaceuticals: Overview on the Production, Purification, and Formulation." Vaccines 9, no. 4 (April 1, 2021): 328. http://dx.doi.org/10.3390/vaccines9040328.

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The advent of biopharmaceuticals in modern medicine brought enormous benefits to the treatment of numerous human diseases and improved the well-being of many people worldwide. First introduced in the market in the early 1980s, the number of approved biopharmaceutical products has been steadily increasing, with therapeutic proteins, antibodies, and their derivatives accounting for most of the generated revenues. The success of pharmaceutical biotechnology is closely linked with remarkable developments in DNA recombinant technology, which has enabled the production of proteins with high specificity. Among promising biopharmaceuticals are interferons, first described by Isaacs and Lindenmann in 1957 and approved for clinical use in humans nearly thirty years later. Interferons are secreted autocrine and paracrine proteins, which by regulating several biochemical pathways have a spectrum of clinical effectiveness against viral infections, malignant diseases, and multiple sclerosis. Given their relevance and sustained market share, this review provides an overview on the evolution of interferon manufacture, comprising their production, purification, and formulation stages. Remarkable developments achieved in the last decades are herein discussed in three main sections: (i) an upstream stage, including genetically engineered genes, vectors, and hosts, and optimization of culture conditions (culture media, induction temperature, type and concentration of inducer, induction regimens, and scale); (ii) a downstream stage, focusing on single- and multiple-step chromatography, and emerging alternatives (e.g., aqueous two-phase systems); and (iii) formulation and delivery, providing an overview of improved bioactivities and extended half-lives and targeted delivery to the site of action. This review ends with an outlook and foreseeable prospects for underdeveloped aspects of biopharma research involving human interferons.

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Shanmugaraj, Balamurugan, Christine Joy I. Bulaon, and Waranyoo Phoolcharoen. "Plant Molecular Farming: A Viable Platform for Recombinant Biopharmaceutical Production." Plants 9, no. 7 (July 4, 2020): 842. http://dx.doi.org/10.3390/plants9070842.

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The demand for recombinant proteins in terms of quality, quantity, and diversity is increasing steadily, which is attracting global attention for the development of new recombinant protein production technologies and the engineering of conventional established expression systems based on bacteria or mammalian cell cultures. Since the advancements of plant genetic engineering in the 1980s, plants have been used for the production of economically valuable, biologically active non-native proteins or biopharmaceuticals, the concept termed as plant molecular farming (PMF). PMF is considered as a cost-effective technology that has grown and advanced tremendously over the past two decades. The development and improvement of the transient expression system has significantly reduced the protein production timeline and greatly improved the protein yield in plants. The major factors that drive the plant-based platform towards potential competitors for the conventional expression system are cost-effectiveness, scalability, flexibility, versatility, and robustness of the system. Many biopharmaceuticals including recombinant vaccine antigens, monoclonal antibodies, and other commercially viable proteins are produced in plants, some of which are in the pre-clinical and clinical pipeline. In this review, we consider the importance of a plant- based production system for recombinant protein production, and its potential to produce biopharmaceuticals is discussed.

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Gerszberg, Aneta, and Katarzyna Hnatuszko-Konka. "Compendium on Food Crop Plants as a Platform for Pharmaceutical Protein Production." International Journal of Molecular Sciences 23, no. 6 (March 17, 2022): 3236. http://dx.doi.org/10.3390/ijms23063236.

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Tremendous advances in crop biotechnology related to the availability of molecular tools and methods developed for transformation and regeneration of specific plant species have been observed. As a consequence, the interest in plant molecular farming aimed at producing the desired therapeutic proteins has significantly increased. Since the middle of the 1980s, recombinant pharmaceuticals have transformed the treatment of many serious diseases and nowadays are used in all branches of medicine. The available systems of the synthesis include wild-type or modified mammalian cells, plants or plant cell cultures, insects, yeast, fungi, or bacteria. Undeniable benefits such as well-characterised breeding conditions, safety, and relatively low costs of production make plants an attractive yet competitive platform for biopharmaceutical production. Some of the vegetable plants that have edible tubers, fruits, leaves, or seeds may be desirable as inexpensive bioreactors because these organs can provide edible vaccines and thus omit the purification step of the final product. Some crucial facts in the development of plant-made pharmaceuticals are presented here in brief. Although crop systems do not require more strictly dedicated optimization of methodologies at any stages of the of biopharmaceutical production process, here we recall the complete framework of such a project, along with theoretical background. Thus, a brief review of the advantages and disadvantages of different systems, the principles for the selection of cis elements for the expression cassettes, and available methods of plant transformation, through to the protein recovery and purification stage, are all presented here. We also outline the achievements in the production of biopharmaceuticals in economically important crop plants and provide examples of their clinical trials and commercialization.

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Samiec, M., and M. Skrzyszowska. "Transgenic mammalian species, generated by somatic cell cloning, in biomedicine, biopharmaceutical industry and human nutrition/dietetics - recent achievements." Polish Journal of Veterinary Sciences 14, no. 2 (May 1, 2011): 317–28. http://dx.doi.org/10.2478/v10181-011-0050-7.

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Transgenic mammalian species, generated by somatic cell cloning, in biomedicine, biopharmaceutical industry and human nutrition/dietetics - recent achievements Somatic cell cloning technology in mammals promotes the multiplication of productively-valuable genetically engineered individuals, and consequently allows also for standardization of transgenic farm animal-derived products, which, in the context of market requirements, will have growing significance. Gene farming is one of the most promising areas in modern biotechnology. The use of live bioreactors for the expression of human genes in the lactating mammary gland of transgenic animals seems to be the most cost-effective method for the production/processing of valuable recombinant therapeutic proteins. Among the transgenic farm livestock species used so far, cattle, goats, sheep, pigs and rabbits are useful candidates for the expression of tens to hundreds of grams of genetically-engineered proteins or xenogeneic biopreparations in the milk. At the beginning of the new millennium, a revolution in the treatment of disease is taking shape due to the emergence of new therapies based on recombinant human proteins. The ever-growing demand for such pharmaceutical or nutriceutical proteins is an important driving force for the development of safe and large-scale production platforms. The aim of this paper is to present an overall survey of the state of the art in investigations which provide the current knowledge for deciphering the possibilities of practical application of the transgenic mammalian species generated by somatic cell cloning in biomedicine, the biopharmaceutical industry, human nutrition/dietetics and agriculture.

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Saraswat, Mayank, Luca Musante, Alessandra Ravidá, Brian Shortt, Barry Byrne, and Harry Holthofer. "Preparative Purification of Recombinant Proteins: Current Status and Future Trends." BioMed Research International 2013 (2013): 1–18. http://dx.doi.org/10.1155/2013/312709.

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Advances in fermentation technologies have resulted in the production of increased yields of proteins of economic, biopharmaceutical, and medicinal importance. Consequently, there is an absolute requirement for the development of rapid, cost-effective methodologies which facilitate the purification of such products in the absence of contaminants, such as superfluous proteins and endotoxins. Here, we provide a comprehensive overview of a selection of key purification methodologies currently being applied in both academic and industrial settings and discuss how innovative and effective protocols such as aqueous two-phase partitioning, membrane chromatography, and high-performance tangential flow filtration may be applied independently of or in conjunction with more traditional protocols for downstream processing applications.

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Hu, Jianwen, Jizhong Han, Haoran Li, Xian Zhang, Lan lan Liu, Fei Chen, and Bin Zeng. "Human Embryonic Kidney 293 Cells: A Vehicle for Biopharmaceutical Manufacturing, Structural Biology, and Electrophysiology." Cells Tissues Organs 205, no. 1 (2018): 1–8. http://dx.doi.org/10.1159/000485501.

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Mammalian cells, e.g., CHO, BHK, HEK293, HT-1080, and NS0 cells, represent important manufacturing platforms in bioengineering. They are widely used for the production of recombinant therapeutic proteins, vaccines, anticancer agents, and other clinically relevant drugs. HEK293 (human embryonic kidney 293) cells and their derived cell lines provide an attractive heterologous system for the development of recombinant proteins or adenovirus productions, not least due to their human-like posttranslational modification of protein molecules to provide the desired biological activity. Secondly, they also exhibit high transfection efficiency yielding high-quality recombinant proteins. They are easy to maintain and express with high fidelity membrane proteins, such as ion channels and transporters, and thus are attractive for structural biology and electrophysiology studies. In this article, we review the literature on HEK293 cells regarding their origins but also stress their advancements into the different cell lines engineered and discuss some significant aspects which make them versatile systems for biopharmaceutical manufacturing, drug screening, structural biology research, and electrophysiology applications.

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Ibrahim, Yousif H.-E. Y., Géza Regdon, Elnazeer I. Hamedelniel, and Tamás Sovány. "Review of recently used techniques and materials to improve the efficiency of orally administered proteins/peptides." DARU Journal of Pharmaceutical Sciences 28, no. 1 (December 6, 2019): 403–16. http://dx.doi.org/10.1007/s40199-019-00316-w.

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Abstract Objectives The main objective of present review is to explore and evaluate the effectiveness of recently developed methods to improve the bioavailability of orally administered biopharmaceutical drugs. Methods A systematic search of sciencedirect, tandfonline and Google Scholar databases based on various sets of keywords was performed. All results were evaluated based on their abstracts, and irrelevant studies were neglected during further evaluation. Results At present, biopharmaceuticals are used as injectable therapies as they are not absorbed adequately from the different routes of drug administration, particularly the oral one. Their insufficient absorption is attributed to their high molecular weight, degradation by proteolytic enzymes, high hydrophilicity and rigidity of the absorptive tissues. From industrial aspect incorporation of enzyme inhibitors (EIs) and permeation enhancers (PEs) and mucoadhesive polymers into conventional dosage forms may be the easiest way of formulation of orally administered macromolecular drugs, but the effectiveness of protection and absorption enhancement here is the most questionable. Conjugation may be problematic from regulatory aspect. Encapsulation into lipid-based vesicles sufficiently protects the incorporated macromolecule and improves intestinal uptake but have considerable stability issues. In contrast, polymeric nanocarriers may provide good stability but provides lower internalization efficacy in comparison with the lipid-based carriers. Conclusion It can be concluded that the combination of the advantages of mucoadhesive polymeric and lid-based carriers in hybrid lipid/polymer nanoparticles may result in improved absorption and might represent a potential means for the oral administration of therapeutic proteins in the near future.

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Hefferon, Kathleen Laura. "Plant virus expression vectors set the stage as production platforms for biopharmaceutical proteins." Virology 433, no. 1 (November 2012): 1–6. http://dx.doi.org/10.1016/j.virol.2012.06.012.

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Zalar, Matja, Hristo L. Svilenov, and Alexander P. Golovanov. "Binding of excipients is a poor predictor for aggregation kinetics of biopharmaceutical proteins." European Journal of Pharmaceutics and Biopharmaceutics 151 (June 2020): 127–36. http://dx.doi.org/10.1016/j.ejpb.2020.04.002.

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Eaton, Leslie C. "Quantitation of residual Escherichia coli DNA in recombinant biopharmaceutical proteins by hybridization analysis." Journal of Pharmaceutical and Biomedical Analysis 7, no. 5 (January 1989): 633–38. http://dx.doi.org/10.1016/0731-7085(89)80230-4.

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Bugelskil, P. J., D. J. Herzykl, S. Rehm, A. G. Harmsen, E. V. Gore, D. M. Williams, B. E. Maleeff, et al. "Preclinical development of keliximab, a Primatized™ anti-CD4 monoclonal antibody, in human CD4 transgenic mice: characterization of the model and safety studies." Human & Experimental Toxicology 19, no. 4 (April 2000): 230–43. http://dx.doi.org/10.1191/096032700678815783.

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The preclinical safety assessment of biopharmaceuticals necessitates that studies be conducted in species in which the products are pharmacologically active. Monoclonal anti-bodies are a promising class ofbiopharmaceuticals for many disease indications; however, by design, these agents tend to have limited species cross-reactivity and tend to only be active in primates. Keliximab is a human-cynomolgus monkey chimeric (Primatized&) monoclonal antibody with specificity for human and chimpanzee CD4. In order to conduct a comprehensive preclinical safety assessment of this antibody to support chronic treatment of rheumatoid arthritis in patients, a human CD4 transgenic mouse was used for chronic and reproductive toxicity studies and for genotoxic studies. In addition, immunotoxicity studies were conducted in these mice with Candida albicans, Pneumo- cystis carinii and B16 melanoma cells to assess the effects of keliximab on host resistance to infection and immunosur-veillance to neoplasia. The results of these studies found keliximab to be well tolerated with the only effects observed being related to its pharmacologic activity on CD4 + T lymphocytes. The use of transgenic mice expressing human proteins provides a useful alternative to studies in chimpan-zees with biopharmaceutical agents having limited species cross-reactivity.

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Keller, G.-A. "Cell Imaging in Drug Discovery and Development." Microscopy and Microanalysis 7, S2 (August 2001): 620–21. http://dx.doi.org/10.1017/s1431927600029172.

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While morphological analysis at both the light and electron microscopical levels has been fully exploited in academic research, its potential in biopharmaceutical industry has only been fully appreciated in recent years. The biotechnology industry has been developing highly specialized therapeutic molecules, particularly proteins and peptides that are directed to specific molecular targets. The pace of the development of new therapeutic molecules is expected to increase dramatically in the close future, as the biopharmaceutical community will capitalize on the new targets identified by the Human Genome Project.An efficient drug discovery and development effort should not only include the characterization of the tissue and intracellular distribution of potential targets, but also establish unambiguously that the therapeutic entities reach their intended targets. It has become apparent in recent years that a comprehensive view of the pharmacological action of drugs must include not only the pharmacokinetic parameters but also the tissue distribution and the cellular localization.

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Cabal, Ace Bryan Sotelo, and Tzong-Yuan Wu. "Recombinant Protein Technology in the Challenging Era of Coronaviruses." Processes 10, no. 5 (May 10, 2022): 946. http://dx.doi.org/10.3390/pr10050946.

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Coronaviruses have caused devastation in both human and animal populations, affecting both health and the economy. Amidst the emergence and re-emergence of coronaviruses, humans need to surmount the health and economic threat of coronaviruses through science and evidence-based approaches. One of these approaches is through biotechnology, particularly the heterologous production of biopharmaceutical proteins. This review article briefly describes the genome, general virion morphology, and key structural proteins of different coronaviruses affecting animals and humans. In addition, this review paper also presents the different systems in recombinant protein technology such as bacteria, yeasts, plants, mammalian cells, and insect/insect cells systems used to express key structural proteins in the development of countermeasures such as diagnostics, prophylaxis, and therapeutics in the challenging era of coronaviruses.

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Bischoff, Rainer, Kees J. Bronsema, Nico C. van de Merbel, Kees J. Bronsema, and Nico C. van de Merbel. "Analysis of biopharmaceutical proteins in biological matrices by LC-MS/MS I. Sample preparation." TrAC Trends in Analytical Chemistry 48 (July 2013): 41–51. http://dx.doi.org/10.1016/j.trac.2012.11.015.

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Bush, David R., Li Zang, Arseniy M. Belov, Alexander R. Ivanov, and Barry L. Karger. "High Resolution CZE-MS Quantitative Characterization of Intact Biopharmaceutical Proteins: Proteoforms of Interferon-β1." Analytical Chemistry 88, no. 2 (December 24, 2015): 1138–46. http://dx.doi.org/10.1021/acs.analchem.5b03218.

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Cardona-Ospina, Jaime A., Juan C. Sepúlveda-Arias, L. Mancilla, and Luis G. Gutierrez-López. "Plant expression systems, a budding way to confront chikungunya and Zika in developing countries?" F1000Research 5 (August 31, 2016): 2121. http://dx.doi.org/10.12688/f1000research.9502.1.

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Plant expression systems could be used as biofactories of heterologous proteins that have the potential to be used with biopharmaceutical aims and vaccine design. This technology is scalable, safe and cost-effective and it has been previously proposed as an option for vaccine and protein pharmaceutical development in developing countries. Here we present a proposal of how plant expression systems could be used to address Zika and chikungunya outbreaks through development of vaccines and rapid diagnostic kits.

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Ibeanu, Nkiruka, Raphael Egbu, Lesley Onyekuru, Hoda Javaheri, Peng Tee Khaw, Gareth R. Williams, Steve Brocchini, and Sahar Awwad. "Injectables and Depots to Prolong Drug Action of Proteins and Peptides." Pharmaceutics 12, no. 10 (October 21, 2020): 999. http://dx.doi.org/10.3390/pharmaceutics12100999.

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Proteins and peptides have emerged in recent years to treat a wide range of multifaceted diseases such as cancer, diabetes and inflammation. The emergence of polypeptides has yielded advancements in the fields of biopharmaceutical production and formulation. Polypeptides often display poor pharmacokinetics, limited permeability across biological barriers, suboptimal biodistribution, and some proclivity for immunogenicity. Frequent administration of polypeptides is generally required to maintain adequate therapeutic levels, which can limit efficacy and compliance while increasing adverse reactions. Many strategies to increase the duration of action of therapeutic polypeptides have been described with many clinical products having been developed. This review describes approaches to optimise polypeptide delivery organised by the commonly used routes of administration. Future innovations in formulation may hold the key to the continued successful development of proteins and peptides with optimal clinical properties.

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Keysberg, Christoph, Oliver Hertel, Louise Schelletter, Tobias Busche, Chiara Sochart, Jörn Kalinowski, Raimund Hoffrogge, Kerstin Otte, and Thomas Noll. "Exploring the molecular content of CHO exosomes during bioprocessing." Applied Microbiology and Biotechnology 105, no. 9 (May 2021): 3673–89. http://dx.doi.org/10.1007/s00253-021-11309-8.

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Abstract In biopharmaceutical production, Chinese hamster ovary (CHO) cells derived from Cricetulus griseus remain the most commonly used host cell for recombinant protein production, especially antibodies. Over the last decade, in-depth multi-omics characterization of these CHO cells provided data for extensive cell line engineering and corresponding increases in productivity. However, exosomes, extracellular vesicles containing proteins and nucleic acids, are barely researched at all in CHO cells. Exosomes have been proven to be a ubiquitous mediator of intercellular communication and are proposed as new biopharmaceutical format for drug delivery, indicator reflecting host cell condition and anti-apoptotic factor in spent media. Here we provide a brief overview of different separation techniques and subsequently perform a proteome and regulatory, non-coding RNA analysis of exosomes, derived from lab-scale bioreactor cultivations of a CHO-K1 cell line, to lay out reference data for further research in the field. Applying bottom-up orbitrap shotgun proteomics and next-generation small RNA sequencing, we detected 1395 proteins, 144 micro RNA (miRNA), and 914 PIWI-interacting RNA (piRNA) species differentially across the phases of a batch cultivation process. The exosomal proteome and RNA data are compared with other extracellular fractions and cell lysate, yielding several significantly exosome-enriched species. Key points • First-time comprehensive protein and miRNA characterization of CHO exosomes. • Isolation protocol and time point of bioprocess strongly affect quality of extracellular vesicles. • CHO-derived exosomes also contain numerous piRNA species of yet unknown function.

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Bracewell, Daniel G., Victoria Smith, Mike Delahaye, and C. Mark Smales. "Analytics of host cell proteins (HCPs): lessons from biopharmaceutical mAb analysis for Gene therapy products." Current Opinion in Biotechnology 71 (October 2021): 98–104. http://dx.doi.org/10.1016/j.copbio.2021.06.026.

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Titchener-Hooker, N. J., P. Dunnill, and M. Hoare. "Micro biochemical engineering to accelerate the design of industrial-scale downstream processes for biopharmaceutical proteins." Biotechnology and Bioengineering 100, no. 3 (2008): 473–87. http://dx.doi.org/10.1002/bit.21788.

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Kim, Nam Ah, Bora Heo, and Seong Hoon Jeong. "Rapid methodology for basal system selection of therapeutic proteins during the early stage biopharmaceutical development." Journal of Pharmaceutical Investigation 50, no. 4 (September 11, 2019): 363–72. http://dx.doi.org/10.1007/s40005-019-00461-z.

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Kotapati, Srikanth, Madhura Deshpande, Aarti Jashnani, Dharam Thakkar, Hongwu Xu, and Gavin Dollinger. "The role of ligand-binding assay and LC–MS in the bioanalysis of complex protein and oligonucleotide therapeutics." Bioanalysis 13, no. 11 (June 2021): 931–54. http://dx.doi.org/10.4155/bio-2021-0009.

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Ligand-binding assay (LBA) and LC–MS have been the preferred bioanalytical techniques for the quantitation and biotransformation assessment of various therapeutic modalities. This review provides an overview of the applications of LBA, LC–MS/MS and LC–HRMS for the bioanalysis of complex protein therapeutics including antibody–drug conjugates, fusion proteins and PEGylated proteins as well as oligonucleotide therapeutics. The strengths and limitations of LBA and LC–MS, along with some guidelines on the choice of appropriate bioanalytical technique(s) for the bioanalysis of these therapeutic modalities are presented. With the discovery of novel and more complex therapeutic modalities, there is an increased need for the biopharmaceutical industry to develop a comprehensive bioanalytical strategy integrating both LBA and LC–MS.

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Van Manen-Brush, Kathleen, Jacob Zeitler, John R. White, Paul Younge, Samantha Willis, and Marisa Jones. "Improving Chinese hamster ovary host cell protein ELISA using Ella®: an automated microfluidic platform." BioTechniques 69, no. 3 (September 2020): 186–92. http://dx.doi.org/10.2144/btn-2020-0074.

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Chinese hamster ovary (CHO) cells are a mammalian cell line used in the production of therapeutic proteins. Host cell proteins (HCPs) are process-related impurities that are derived from the host cell expression system. During biopharmaceutical drug development, removal of HCPs is required. Enzyme-linked immunosorbent assay (ELISA) is a common technique to quantitate HCPs, but is a labor-intensive process that takes up to 7 h. Ella® is an automated instrument that utilizes microfluidics and glass nanoreactors to quantitate HCPs in 75 min using similar ELISA reagents. The antibodies and antigens are captured on three distinct glass nanoreactors, resulting in sensitive reproducible data. Our results indicate that Ella quantitates CHO HCPs with precision, accuracy, sensitivity and trends comparable with our traditional CHO HCP ELISA.

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Zhong, Xiaotian, and Jill F. Wright. "Biological Insights into Therapeutic Protein Modifications throughout Trafficking and Their Biopharmaceutical Applications." International Journal of Cell Biology 2013 (2013): 1–19. http://dx.doi.org/10.1155/2013/273086.

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Over the lifespan of therapeutic proteins, from the point of biosynthesis to the complete clearance from tested subjects, they undergo various biological modifications. Therapeutic influences and molecular mechanisms of these modifications have been well appreciated for some while remained less understood for many. This paper has classified these modifications into multiple categories, according to their processing locations and enzymatic involvement during the trafficking events. It also focuses on the underlying mechanisms and structural-functional relationship between modifications and therapeutic properties. In addition, recent advances in protein engineering, cell line engineering, and process engineering, by exploring these complex cellular processes, are discussed and summarized, for improving functional characteristics and attributes of protein-based biopharmaceutical products.

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Dewi, Kartika Sari, and Asrul Muhamad Fuad. "Improving the Expression of Human Granulocyte Colony Stimulating Factor in Escherichia coli by Reducing the GC-content and Increasing mRNA Folding Free Energy at 5’-Terminal End." Advanced Pharmaceutical Bulletin 10, no. 4 (August 9, 2020): 610–16. http://dx.doi.org/10.34172/apb.2020.073.

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Purpose : Strategy for improving the production of biopharmaceutical protein continues to develop due to increasing market demand. Human granulocyte colony stimulating factor (hGCSF) is one of biopharmaceutical proteins that has many applications, and easily produced in Escherichia coli expression system. Previous studies reported that codon usage, rare codon, mRNA folding and GC-content at 5’-terminal end were crucial for protein production in E. coli. In the present study, the effect of reducing the GC-content and increasing the mRNA folding free energy at the 5’-terminal end on the expression level of hG-CSF proteins was investigated. Methods: Synonymous codon substitutions were performed to generate mutant variants of open reading frame (ORF) with lower GC-content at 5’-terminal ends. Oligoanalyzer tool was used to calculate the GC content of eight codons sequence after ATG. Whereas, mRNA folding free energy was predicted using KineFold and RNAfold tools. The template DNA was amplified using three variant forward primers and one same reverse primer. Those DNA fragments were individually cloned into pJexpress414 expression vector and were confirmed using restriction and DNA sequencing analyses. The confirmed constructs were transformed into E. coli NiCo21(DE3) host cells and the recombinant protein was expressed using IPTG-induction. Total protein obtained were characterized using SDS-PAGE, Western blot and ImageJ software analyses. Results: The result showed that the mutant variant with lower GC-content and higher mRNA folding free energy near the translation initiation region (TIR) could produce a higher amount of hG-CSF proteins compared to the original gene sequence. Conclusion: This study emphasized the important role of the nucleotide composition immediately downstream the start codon to achieve high-yield protein product on heterologous expression in E. coli.

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Benedé, Sara, and Elena Molina. "Chicken Egg Proteins and Derived Peptides with Antioxidant Properties." Foods 9, no. 6 (June 3, 2020): 735. http://dx.doi.org/10.3390/foods9060735.

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In addition to their high nutritional value, some chicken egg proteins and derivatives such as protein hydrolysates, peptides and amino acids show antioxidant properties which make them prominent candidates for the development of functional foods, drawing attention to both the food and biopharmaceutical industries. This review summarizes current knowledge on antioxidant activity of chicken egg proteins and their derived peptides. Some egg proteins such as ovalbumin, ovotransferrin and lysozyme from egg white or phosvitin from yolk have shown antioxidant properties, although derived peptides have higher bioactive potential. The main process for obtaining egg bioactive peptides is enzymatic hydrolysis of its proteins using enzymes and/or processing technologies such as heating, sonication or high-intensity-pulsed electric field. Different in vitro assays such as determination of reducing power, DPPH and ABTS radical-scavenging activity tests or oxygen radical absorbance capacity assay have been used to evaluate the diverse antioxidant mechanisms of proteins and peptides. Similarly, different cell lines and animal models including zebrafish, mice and rats have also been used. In summary, this review collects all the knowledge described so far regarding egg proteins and derived peptides with antioxidant functions.

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Inouye, Masaharu, and Thierry Burnouf. "The Role of Nanofiltration in the Pathogen Safety of Biologicals: An Update." Current Nanoscience 16, no. 3 (April 2, 2020): 413–24. http://dx.doi.org/10.2174/1573413715666190328223130.

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Nanofiltration technology to remove possible pathogenic viruses during biopharmaceutical manufacturing was introduced in the biopharmaceutical industry in 1989. The very first industrial implementation took place in the early 1990s, through commercial manufacturing processes of plasma- derived medical products. Then it was applied to recombinant protein medical products, including monoclonal antibodies. In the first review published in 2005 in this journal, the technology was already considered promising and was much welcomed by the industry, but it was still a relatively emerging technology at that time, and many questions were raised about its robustness as a reliable virus-removal tool. We conducted a review to update the published information (SCI journals and suppliers’ documentation) existing on the use of nanofiltration as an industrial process for removing viruses from various biologicals. After almost a decade from the previous review, nanofiltration has established itself as a routine production step in most biopharmaceutical manufacturing. It has become one of the essential manufacturing processes used to assure safety against viral contamination. The technology is applied to manufacturing processes of various biologicals (human plasma products and complex recombinant proteins, such as coagulation factors and monoclonal antibodies made from mammalian cells). Many biologicals that undergo nanofiltration are licensed by regulatory authorities, which illustrates that nanofiltration is recognized as a robust and safe virus-removal method. No adverse events related to the use of nanofiltration have been recorded. New trends in nanofiltration technology continue to appear. As was identified during its introduction to the market and predicted in the previous review, nanofiltration has achieved major technical breakthroughs for ensuring the safety of biologicals, particularly human plasma-derived products, against viruses.

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Gebauer, Michaela, and Arne Skerra. "Engineered Protein Scaffolds as Next-Generation Therapeutics." Annual Review of Pharmacology and Toxicology 60, no. 1 (January 6, 2020): 391–415. http://dx.doi.org/10.1146/annurev-pharmtox-010818-021118.

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The concept of engineering robust protein scaffolds for novel binding functions emerged 20 years ago, one decade after the advent of recombinant antibody technology. Early examples were the Affibody, Monobody (Adnectin), and Anticalin proteins, which were derived from fragments of streptococcal protein A, from the tenth type III domain of human fibronectin, and from natural lipocalin proteins, respectively. Since then, this concept has expanded considerably, including many other protein templates. In fact, engineered protein scaffolds with useful binding specificities, mostly directed against targets of biomedical relevance, constitute an area of active research today, which has yielded versatile reagents as laboratory tools. However, despite strong interest from basic science, only a handful of those protein scaffolds have undergone biopharmaceutical development up to the clinical stage. This includes the abovementioned pioneering examples as well as designed ankyrin repeat proteins (DARPins). Here we review the current state and clinical validation of these next-generation therapeutics.

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Thoring, Zemella, Wüstenhagen, and Kubick. "Accelerating the Production of Druggable Targets: Eukaryotic Cell-Free Systems Come into Focus." Methods and Protocols 2, no. 2 (April 16, 2019): 30. http://dx.doi.org/10.3390/mps2020030.

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In the biopharmaceutical pipeline, protein expression systems are of high importance not only for the production of biotherapeutics but also for the discovery of novel drugs. The vast majority of drug targets are proteins, which need to be characterized and validated prior to the screening of potential hit components and molecules. A broad range of protein expression systems is currently available, mostly based on cellular organisms of prokaryotic and eukaryotic origin. Prokaryotic cell-free systems are often the system of choice for drug target protein production due to the simple generation of expression hosts and low cost of preparation. Limitations in the production of complex mammalian proteins appear due to inefficient protein folding and posttranslational modifications. Alternative protein production systems, so-called eukaryotic cell-free protein synthesis systems based on eukaryotic cell-lysates, close the gap between a fast protein generation system and a high quality of complex mammalian proteins. In this study, we show the production of druggable target proteins in eukaryotic cell-free systems. Functional characterization studies demonstrate the bioactivity of the proteins and underline the potential for eukaryotic cell-free systems to significantly improve drug development pipelines.

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Gagnon, Pete, and Tsutomu Arakawa. "Editorial [Hot Topic: Aggregation Detection and Removal Biopharmaceutical Proteins (Guest Editors: Pete Gagnon & Tsutomu Arakawa)]." Current Pharmaceutical Biotechnology 10, no. 4 (June 1, 2009): 347. http://dx.doi.org/10.2174/138920109788488923.

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Hopfgartner, Gérard, Antoine Lesur, and Emmanuel Varesio. "Analysis of biopharmaceutical proteins in biological matrices by LC-MS/MS II. LC-MS/MS analysis." TrAC Trends in Analytical Chemistry 48 (July 2013): 52–61. http://dx.doi.org/10.1016/j.trac.2013.03.008.

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Kouwen, Thijs R. H. M., Jean-Yves F. Dubois, Roland Freudl, Wim J. Quax, and Jan Maarten van Dijl. "Modulation of Thiol-Disulfide Oxidoreductases for Increased Production of Disulfide-Bond-Containing Proteins in Bacillus subtilis." Applied and Environmental Microbiology 74, no. 24 (October 24, 2008): 7536–45. http://dx.doi.org/10.1128/aem.00894-08.

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ABSTRACT Disulfide bonds are important for the correct folding, structural integrity, and activity of many biotechnologically relevant proteins. For synthesis and subsequent secretion of these proteins in bacteria, such as the well-known “cell factory” Bacillus subtilis, it is often the correct formation of disulfide bonds that is the greatest bottleneck. Degradation of inefficiently or incorrectly oxidized proteins and the requirement for costly and time-consuming reduction and oxidation steps in the downstream processing of the proteins still are major limitations for full exploitation of B. subtilis for biopharmaceutical production. Therefore, the present study was aimed at developing a novel in vivo strategy for improved production of secreted disulfide-bond-containing proteins. Three approaches were tested: depletion of the major cytoplasmic reductase TrxA; introduction of the heterologous oxidase DsbA from Staphylococcus carnosus; and addition of redox-active compounds to the growth medium. As shown using the disulfide-bond-containing molecule Escherichia coli PhoA as a model protein, combined use of these three approaches resulted in secretion of amounts of active PhoA that were ∼3.5-fold larger than the amounts secreted by the parental strain B. subtilis 168. Our findings indicate that Bacillus strains with improved oxidizing properties can be engineered for biotechnological production of heterologous high-value proteins containing disulfide bonds.

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Journal articles on the topic 'Biopharmaceutical proteins'

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