Production of Monoclonal Antibodies in Serum-free Media

Manna, Laura; Febo, Tiziana Di; Armillotta, Gisella; Luciani, Mirella; Ciarelli, Antonella; Salini, Romolo; Ventura, Mauro Di

doi:10.1089/mab.2015.0004

Cited by 9 publications

(6 citation statements)

References 21 publications

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“…Serum free medium has also been successfully used in high cell density bioreactors such as hollow fiber systems [ 18 ], packed bed reactors [ 19 ] and wave bioreactors [ 20 ] for the cultivation of mammalian cells. There is a considerable progress in cell culture techniques using serum free media, that supports the growth of hybridoma cells [ 21 ] and CHO cell lines [ 22 , 23 ] for therapeutic protein production. To reduce the cost of production and to increase the output in therapeutic protein production, high-density culture (≤10 7 cells/ml) systems have been introduced.…”

Section: Track To the Emergence Of High-density Cell Culture Systemsmentioning

confidence: 99%

Trends in Monoclonal Antibody Production Using Various Bioreactor Syst

Jyothilekshmi

Jayaprakash

2021

J. Microbiol. Biotechnol.

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Monoclonal antibodies are widely used as diagnostic reagents and for therapeutic purposes, and their demand is increasing extensively. To produce these proteins in sufficient quantities for commercial use, it is necessary to raise the output by scaling up the production processes. This review describes recent trends in high-density cell culture systems established for monoclonal antibody production that are excellent methods to scale up from the lab-scale cell culture. Among the reactors, hollow fiber bioreactors contribute to a major part of high-density cell culture as they can provide a tremendous amount of surface area in a small volume for cell growth. As an alternative to hollow fiber reactors, a novel disposable bioreactor has been developed, which consists of a polymer-based supermacroporous material, cryogel, as a matrix for cell growth. Packed bed systems and disposable wave bioreactors have also been introduced for high cell density culture. These developments in high-density cell culture systems have led to the monoclonal antibody production in an economically favourable manner and made monoclonal antibodies one of the dominant therapeutic and diagnostic proteins in biopharmaceutical industry.

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Section: Track To the Emergence Of High-density Cell Culture Systemsmentioning

confidence: 99%

Trends in Monoclonal Antibody Production Using Various Bioreactor Syst

Jyothilekshmi

Jayaprakash

2021

J. Microbiol. Biotechnol.

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“…Since the environment may have an influence on serum content, the risk of pathogens contamination must be considered too. Moreover, serum supplementation in a culture medium during the production of MAb gives impurities and causes difficulties in the purification process [5][6][7][8][9][10].…”

Section: Introductionmentioning

confidence: 99%

The Effect of Indonesian Royal Jelly Supplementation on the Growth of Hybridoma and Its Monoclonal Antibody Production

Ningsih¹,

Widayanti²,

Sahlan³

et al. 2023

Proceedings of the 1st International Conference for Health Research – BRIN (ICHR 2022)

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Monoclonal antibody (mAb) is the material that is used widely for biological experiments, therapeutic, and diagnosis. The production of monoclonal antibodies by in vitro method is still challenging, and it takes a high cost due to the use of animal-derived serum as the main growth-promoting factor. The previous study reported royal jelly (RJ) from Apis mellifera as a potential proliferation factor in mammalian cell culture. Hence, this study was performed to evaluate the effect of Indonesian RJ supplementation in culture medium on hybridoma growth and its MAb production. The research was conducted by true experimental methods using hybridoma clone SF2 and RF10. These clones were cultured in RPMI-1640 medium supplemented with 3% FBS and 1% Penicillin-Streptomycin as the initial culture medium. The number of viable cells was counted by haemocytometer on day 8 after gradual treatment with or without the addition of 1% RJ. MTT assay was performed to evaluate the cell viability. In addition, the level of secreted MAb in the culture medium was measured by indirect enzyme-linked immunosorbent assay (ELISA). All results were analyzed statistically. The results show that the addition of 1% RJ into the culture medium has decreased the cell viability compared to the control. Thus, cell viability reduction may influence the level of MAb produced by hybridoma clone SF2 and RF10. The negative result given in this experiment may have been caused by an unsuitable concentration of RJ used in this experiment. Also, a substance included in the crude RJ probably causes cytotoxicity in the cells.

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“…Serum-free media are commonly used in the production of monoclonal antibodies, vaccines, and other biological products (Lohr et al 2010;Peschel et al 2013;Rourou et al 2014;Genzel 2015;Manna et al 2015;Zhang et al 2017). Serum-free platforms have been described for viral vaccine production, with examples including the use of rabies virus in Vero, BHK-21 and HEK-293 cells (Merten et al 1994(Merten et al , 1999Perrin et al 1995;Kallel et al 2002;Frazatti-Gallina et al 2004;Peschel et al 2013;Fontana et al 2015); foot-and-mouth disease virus in BHK-21 cells (Barteling and Vreeswijk 1991); influenza virus in MDCK cells and HEK293SF-3F6 cells (Brands et al 1999;Petiot et al 2011;Huang et al 2015), and poliovirus in Vero cells (Cinatl et al 1993;Merten et al 1994).…”

Section: Introductionmentioning

confidence: 99%

Semliki Forest Virus replicon particles production in serum-free medium BHK-21 cell cultures and their use to express different proteins

et al. 2019

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The production of biopharmaceuticals as vaccines in serum-free media results in reduced risk of contamination and simpler downstream processing. The production of enveloped viruses and viral vectors such as Semliki Forest Virus (SFV) typically requires lipids that are provided by supplementation with animal serum, so production under serum-free conditions is challenging. In this work, the capacity to deliver genetic material of SFV-viral replicon particles (SFV-VRPs) produced in BHK-21 cells adapted to serum-free medium (BHK/SFM) was evaluated. Three transgenes were evaluated: GFP used as a model protein, while hepatitis C virus nonstructural protein 3 protease domain (HCV-NS3p) and rabies virus glycoprotein (RVGP) were selected based on their distinct nature (enzyme and glycoprotein, respectively). BHK/SFM cells produced a sevenfold higher number of SFV-VRPs, as determined by qRT-PCR. These particles showed similar capacities of infecting BHK/FBS or BHK/SFM cells. GFP expression was evaluated by flow cytometry, HCV-NS3p activity by enzymatic assay, and RVGP expression by ELISA and Western Blot. Expression analysis revealed higher levels of GFP and HCV-NS3p in BHK/SFM, while the levels of RVGP were similar for BHK/SFM and BHK/ FBS. In conclusion, the BHK/SFM cells showed increased SFV-VRP production yields, without affecting vector infectivity or heterologous gene expression, hence validating the use of BHK/SFM for industrial applications. Keywords BHK-21 cells Á Hepatitis C virus nonstructural protein 3 protease domain Á Rabies virus glycoprotein Á Serum-free medium Á Semliki Forest Virus Á Viral replicon particles Abbreviations SFV-VRPs Viral replicon particles of Semliki Forest Virus BHK/FBS BHK-21 cells grown in DMEM supplemented with 10% of fetal bovine serum Carlos Augusto Pereira-Died October 27th, 2017.

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Production of Monoclonal Antibodies in Serum-free Media

Cited by 9 publications

References 21 publications

Trends in Monoclonal Antibody Production Using Various Bioreactor Syst

Trends in Monoclonal Antibody Production Using Various Bioreactor Syst

The Effect of Indonesian Royal Jelly Supplementation on the Growth of Hybridoma and Its Monoclonal Antibody Production

Semliki Forest Virus replicon particles production in serum-free medium BHK-21 cell cultures and their use to express different proteins

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