Papers by Adolfo Castillo
BMC Biotechnology, Nov 22, 2011
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Biotecnología Aplicada, 1994
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Biotecnología Aplicada, Dec 1, 2013
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BMC Infectious Diseases, 2019
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Frontiers in Immunology, 2017
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Vaccine, 2014
The development of safe, effective, and affordable vaccines has become a global effort due to its... more The development of safe, effective, and affordable vaccines has become a global effort due to its vast impact on overall world health conditions. A brief overview of vaccine characterization techniques, especially in the area of high-resolution mass spectrometry, is presented. It is highly conceivable that the proper use of advanced technologies such as high-resolution mass spectrometry, along with the appropriate chemical and physical property evaluations, will yield tremendous in-depth scientific understanding for the characterization of vaccines in various stages of vaccine development. This work presents the physicochemical and biological characterization of cancer vaccine Racotumomab/alumina, a murine anti-idiotypic antibody that mimics N-glycolyl-GM3 gangliosides. This antibody has been tested as an anti-idiotypic cancer vaccine, adjuvated in Al(OH)3, in several clinical trials for melanoma, breast, and lung cancer. Racotumomab was obtained from ascites fluid, transferred to fermentation in stirred tank at 10 L and followed to a scale up to 41 L. The mass spectrometry was used for the determination of intact molecule, light and heavy chains masses; amino acids sequence analysis, N- and C-terminal, glycosylation and posttranslational modifications. Also we used the DLS for the size distribution and zeta potential analysis. The biological analyses were performed in mice and chickens. We observed differences in glycosylation pattern, charge heterogeneity and structural stability between in vivo-produced and bioreactor-obtained Racotumomab products. Interestingly, these modifications had no significant impact on the immune responses elicited in two different animal models. We are demonstrated that this approach could potentially be more efficient and effective for supporting vaccine research and development.
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Animal Cell Technology: Products from Cells, Cells as Products
The use of mammalian cell lines for expression of recombinant proteins with potential therapeutic... more The use of mammalian cell lines for expression of recombinant proteins with potential therapeutic use has become increasingly prevalent.
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New Developments and New Applications in Animal Cell Technology
NSO transfected myeloma producing an anti-EGF receptor humanized MAb was cultivated in hollow fib... more NSO transfected myeloma producing an anti-EGF receptor humanized MAb was cultivated in hollow fiber bioreactors up to production scale (Acusyst-P3/X). Its production rate and metabolic uptakes were compared with a hybridoma, that secreted the murine variant of this product also produced in hollow fiber. The control strategy of the fermentation is discussed. The purification scheme, mainly based on affinity and
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Journal of Chromatography B, 2009
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Cytotechnology, 2011
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World Journal of Vaccines, 2014
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Semicontinuous cultures are a powerful tool for cell characterization. However, it is hard to be ... more Semicontinuous cultures are a powerful tool for cell characterization. However, it is hard to be established at small scale due to the far more complex experimental configuration required. In this work, the use of semicontinuous culture in spinner flasks was explored with daily feeds and extractions, considering that metabolic and growth rates are lower in animal cells under continuous culture regimes. Relatively stabilized conditions were established, characterized by small fluctuations in the concentration of the different metabolic species (glucose, lactate, etc.) at the dilution rates evaluated, and the kinetic parameters (μ, qp, qGlu) were calculated as a function of the dilution rate for each phase. Results obtained were equivalent to those expected for a continuous culture, the semicontinuous operation mode being a much simpler variant. Up to now there are only scarce reports on using semicontinuous cultures as a feasible alternative to simulate the kinetic behavior at statio...
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Semicontinuous cultures are a powerful tool for cell characterization. However, it is hard to be ... more Semicontinuous cultures are a powerful tool for cell characterization. However, it is hard to be established at small scale due to the far more complex experimental configuration required. In this work, the use of semicontinuous culture in spinner flasks was explored with daily feeds and extractions, considering that metabolic and growth rates are lower in animal cells under continuous culture regimes. Relatively stabilized conditions were established, characterized by small fluctuations in the concentration of the different metabolic species (glucose, lactate, etc.) at the dilution rates evaluated, and the kinetic parameters (μ, qp, qGlu) were calculated as a function of the dilution rate for each phase. Results obtained were equivalent to those expected for a continuous culture, the semicontinuous operation mode being a much simpler variant. Up to now there are only scarce reports on using semicontinuous cultures as a feasible alternative to simulate the kinetic behavior at statio...
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Biotecnología Aplicada, 2013
Valorization of knowledge generated by the fast advance of science and its marketing has led to t... more Valorization of knowledge generated by the fast advance of science and its marketing has led to the so-called “knowledge-based economy” and as a consequence high-technology (HiTech) companies have emerged. Those based in Biotechnology have some specifi c features compared to other HiTech sectors. This paper provides a model to structure the economic management of HiTech Biotechnology organizations in Cuba, considering the national economy and the experience accumulated by several institutions from the Scientifi c Biotech Pole in Western Havana. The model is based on some concepts and proposals, and it addresses key elements, such as: insurance of adequate funding levels for R&D activities and technology replacement, fl exible import and export management, the ability to get into very competitive markets and preservation of a highly qualifi ed workforce. It also demonstrates the feasibility to establish this kind of enterprises in the context and regulations already existing in a non...
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Semicontinuous cultures as a tool for characterization of murine hybridoma 1E10 cells. Semicontin... more Semicontinuous cultures as a tool for characterization of murine hybridoma 1E10 cells. Semicontinuous cultures are a powerful tool for cell characterization. However, it is hard to be established at small scale due to the far more complex experimental configuration required. In this work, the use of semicontinuous culture in spinner flasks was explored with daily feeds and extractions, considering that metabolic and growth rates are lower in animal cells under continuous culture regimes. Relatively stabilized conditions were established, characterized by small fluctuations in the concentration of the different metabolic species (glucose, lactate, etc.) at the dilution rates evaluated, and the kinetic parameters (m, qp, qGlu) were calculated as a function of the dilution rate for each phase. Results obtained were equivalent to those expected for a continuous culture, the semicontinuous operation mode being a much simpler variant. Up to now there are only scarce reports on using semic...
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The extensive and labor-intensive process of selection and adaptation of a cell line to culture m... more The extensive and labor-intensive process of selection and adaptation of a cell line to culture medium is the biggest limitation of recombinant protein production in mammalian cell culture, due to a very low frequency of the desired clones within a heterogeneously-distributed population of cells. In this study, the kinetic parameters of different clones of a recombinant NS0 myeloma cell line, obtained by serial cloning and adaptation to protein-free medium and secreting the hR3 IgG1 immunoglobulin, were compared. The physico-chemical parameters, biological activity and antibody affinity constant of the antibody produced was determined for each clone. Two culture systems were used to evaluate the effect of culture conditions on antibody structure and functional heterogeneity: adhesion cell cultures in hollow fiber bioreactors and suspension cultures in spinner flasks. A distinct impact of adaptation and selection processes was evidenced on the microheterogeneity of the cell populatio...
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New Developments and New Applications in Animal Cell Technology, 2002
The recombinant cell line R3/T16 expresses a “reshaped” Mab against epidermal growth factor recep... more The recombinant cell line R3/T16 expresses a “reshaped” Mab against epidermal growth factor receptor (EGF-R) in NSO myeloma. In this study the ability of two culture media to mantain good levels of growth and production of this cell line under serum reduced conditions have been compared and the effect of supplementation of aminoacids and vitamins at 1% of fetal calf
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Papers by Adolfo Castillo