Biopharmaceuticals from microorganisms: from production to purification

Jozala, Ângela Faustino; Geraldes, Danilo Costa; Tundisi, Louise Lacalendola; Feitosa, Valker de Araújo; Breyer, Carlos Alexandre; Cardoso, Samuel Leite; Mazzola, Priscila Gava; Nascimento, Laura de Oliveira; Rangel-Yagui, Carlota de Oliveira; Magalhães, Pérola Oliveira; Oliveira, Marcos Antônio de; Pessoa, Adalberto

doi:10.1016/j.bjm.2016.10.007

Cited by 135 publications

(109 citation statements)

References 73 publications

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“…Heterologous protein expression is a standard workflow common in numerous fields of biology and E. coli is the workhorse microbe for routine protein production in academia and industry. [1][2][3][4] E. coli based processes are used for the production of over 30% of protein based drugs that are on the market today, [5][6][7] and pET based expression in E. coli strain BL21(DE3) and its derivatives is a mainstay of heterologous expression in many labs.…”

Section: Introductionmentioning

confidence: 99%

Improved, scalable, two-stage, autoinduction of recombinant protein expression in E. coli utilizing phosphate depletion

Menacho‐Melgar

Moreb

et al. 2019

Preprint

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We report the improved production of recombinant proteins in E. coli , reliant on tightly controlled autoinduction, triggered by phosphate depletion in stationary phase. The method, reliant on engineered strains and plasmids, enables improved protein expression across scales.Expression levels using this approach have reached as high as 55% of total cellular protein.Initial use of the method in instrumented fed batch fermentations enables cell densities of 10 grams dry cell weight (gCDW) per liter and protein titers up to 2.7+/-0.2 g/L (270 mg/gCDW).The process has also been adapted to an optimized autoinduction media, enabling routine batch production at culture volumes of 20 L (384 well plates), 100 L (96 well plates), 20 mL and 100 mL. In batch cultures, cells densities routinely reach~5-7 gCDW per liter, offering protein titers above 2 g/L. The methodology has been validated with a set of diverse heterologous proteins and is of general use for the facile optimization of routine protein expression from high throughput screens to fed-batch fermentation Highlights :• Stationary phase protein expression results in high titers.• Autoinduction by phosphate depletion enables protein titers from 2-3 g/L.• Autoinduction has been validated from 384 well plates to instrumented bioreactors.

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Section: Introductionmentioning

confidence: 99%

Improved, scalable, two-stage, autoinduction of recombinant protein expression in E. coli utilizing phosphate depletion

Menacho‐Melgar

Moreb

et al. 2019

Preprint

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“…human asparaginase (rhASP) is an important and bestselling chemotherapeutic drug in the world. [1] It (EC 3.5.1.1) is an effective antileukemic drug that catalysis hydrolysis of asparagine into L-aspartic acid and ammonium. This enzyme occurs in microbes, plants, and animals.…”

Section: R Ecombinantmentioning

confidence: 99%

Untitled

Ronda¹

2019

IJGP

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Aim: Recombinant human asparaginase (rhASP) from Escherichia coli is an important therapeutic enzyme used in the treatment of malignant cancers. Due to such a pivotal role, rhASP production in E. coli has drawn great attention of the biopharmaceutical market commercially. The present work aims at the optimization of fermentation medium for the production of rhASP in E. coli. Materials and Methods: The rhASP yield is optimized using sequential optimization designs comprising one variable at a time, Taguchi design, and central composite designs (CCDs). Taguchi design is used to select the effective variables such as soytone, sodium pyruvate, trace element solution, vitamin solution, and yeast extract, which are further optimized by CCD under response surface methodology. Results and Discussion: The CCD design developed a quadratic model with high adequacy for the prediction of rhASP yield with a statistically significant response (R 2 = 97.49% and P < 0.0001) toward the variables. Conclusion: The CCD results showed that the maximum rhASP yield of 38.4387 µg/mL was achieved with the optimized concentrations of media components comprising 9.0 g/L of soytone, 7.5 g/L of sodium pyruvate, 12.5 mL/L of trace element solution, 12.5 mL/L of vitamin solution, and 9.0 g/L of yeast extract.

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“…However, as a measure to increase reproducibility and minimize risks, the trend in the biopharmaceutical industry is now towards the use of animal‐free components . The DSPs, comprise all steps used to recover and purify the product from the cell culture . Both USPs and DSPs are essential in the manufacturing of biopharmaceuticals and should be mutually considered when assessing these type of products.…”

Section: Introductionmentioning

confidence: 99%

“…18,19 The DSPs, comprise all steps used to recover and purify the product from the cell culture. 20,21 Both USPs and DSPs are essential in the manufacturing of biopharmaceuticals and should be mutually considered when assessing these type of products.…”

Section: Introductionmentioning

confidence: 99%

Environmental sustainability assessment of the manufacturing process of a biological active pharmaceutical ingredient

Gamiz

Soete

Heirman

et al. 2019

J of Chemical Tech & Biotech

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BACKGROUND Although the biopharmaceutical market grows rapidly, the environmental impacts of biopharmaceutical manufacturing have been hardly evaluated. The aim of the study is to comprehensively assess the environmental sustainability of a biologically‐produced active pharmaceutical ingredient (API) manufacturing process. For this purpose, an exergy analysis and an (exergetic) life cycle assessment were performed using primary (foreground system) and modeled data (background system). Infliximab, a monoclonal antibody, was considered. RESULTS The fermentation and its raw materials' supply chain was determined to have the highest environmental impact, since the culture media requires chemicals and complex compounds such as animal‐derived materials (ADMs). The total impact per 100 mg of API produced ranged between 18.6 and 101.8 MJex and 2.0–3.1 kg CO2‐eq, mainly as a function of the ADMs' source. A comparison of infliximab and ustekinumab, a biopharmaceutical produced without ADMs, showed that eliminating these materials can reduce the cumulative resource consumption of monoclonal antibody manufacturing by up to 7.5 times. The fermentation is the process taking the longest and the heating, ventilation and air conditioning system which support fermentation areas consumes approximately 75% of all plant electricity use. CONCLUSIONS Culture media nutrients and cleanroom requirements are driving factors in the environmental impact of biological API manufacturing. Obtaining more precise data of the bioprocesses behind these nutrients' supply chain is crucial to corroborating this study's results. © 2019 Society of Chemical Industry

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Biopharmaceuticals from microorganisms: from production to purification

Cited by 135 publications

References 73 publications

Improved, scalable, two-stage, autoinduction of recombinant protein expression in E. coli utilizing phosphate depletion

Improved, scalable, two-stage, autoinduction of recombinant protein expression in E. coli utilizing phosphate depletion

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Environmental sustainability assessment of the manufacturing process of a biological active pharmaceutical ingredient

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