A supervision and control tool based on artificial intelligence for high cell density cultivations

Horta, Antonio Carlos Luperni; Silva, A. J.; Sargo, Cíntia Regina; Velez, Ana; Gonzaga, Miguel; Giordano, Roberto de Campos; Gonçalves, Viviane Maimoni; Zangirolami, Teresa Cristina

doi:10.1590/0104-6632.20140312s00002304

Cited by 21 publications

(25 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The protein was produced and purified using previously published methods with slight modifications Horta et al, 2012;Horta et al, 2014). The production of recombinant PspA4Pro was performed in 5 L bioreactors using; fed-batch cultivation with defined medium containing glycerol as carbon source and lactose as inducer (Horta et al, 2012) or batch cultivation with complex medium containing glucose, glycerol and lactose for auto-induction (Horta et al, 2014). The purification method consisted of cell disruption in a continuous high pressure homogenizer, precipitation of the homogenate with cetyltrimethylammonium bromide, pellet removal by centrifugation, anion exchange chromatography in Q-Sepharose, cryoprecipitation and cation exchange chromatography in SP-Sepharose .…”

Section: Production and Purification Of Pspa4promentioning

confidence: 99%

Pulmonary dry powder vaccine of pneumococcal antigen loaded nanoparticles

Kunda

Alfagih

Miyaji

et al. 2015

International Journal of Pharmaceutics

View full text Add to dashboard Cite

Pneumonia, caused by Streptococcus pneumoniae, mainly affects the immune compromised, the very young and the old, and remains one of the leading causes of death.A steady rise in disease numbers from non-vaccine serotypes necessitates a new vaccine formulation that ideally has better antigen stability and integrity, does not require cold-chain and can be delivered non-invasively. In this study, a dry powder vaccine containing an important antigen of S. pneumoniae, pneumococcal surface protein A (PspA) that has shown cross-reactivity amongst serotypes to be delivered via the pulmonary route has been formulated. The formulation contains the antigen PspA adsorbed onto the surface of polymeric nanoparticles encapsulated in L-leucine microparticles that can be loaded into capsules and delivered via an inhaler. We have successfully synthesized particles of ~150 nm and achieved ~ 20 µg of PspA adsorption per mg of NPs. In addition, the spray-dried powders displayed a FPF of 74.31±1.32% and MMAD of 1.70±0.03 μm suggesting a broncho-alveolar lung deposition facilitating the uptake of the nanoparticles by dendritic cells. Also, the PspA released from the dry powders maintained antigen stability (SDS-PAGE), integrity (Circular dichroism) and activity (lactoferrin binding assay). Moreover, the released antigen also maintained its antigenicity as determined by ELISA.

show abstract

Section: Production and Purification Of Pspa4promentioning

confidence: 99%

Pulmonary dry powder vaccine of pneumococcal antigen loaded nanoparticles

Kunda

Alfagih

Miyaji

et al. 2015

International Journal of Pharmaceutics

View full text Add to dashboard Cite

show abstract

“…Online data were used to estimate cell growth rate. The supervisory system SuperSys_HCDC R (Horta et al 2011Horta et al 2012), based on LabView (National Instruments), was used to monitor and control the experimental apparatus.…”

Section: Methodsmentioning

confidence: 99%

“…In Experiments #3 and #4, both C 1 and C 2 were continuously retuned at each 10 min using the values of μ obtained online from the permittivity measurements provided by the biomass sensor (Horta et al 2011). Thus, for Experiment #3 and #4, Equations 1 and 2 are no more based on a phenomenological reasoning that imposes a fixed value for μ.…”

Section: Methodsmentioning

confidence: 99%

Non-conventional induction strategies for production of subunit swine erysipelas vaccine antigen in rE. coli fed-batch cultures

et al. 2013

View full text Add to dashboard Cite

In spite of the large number of reports on fed-batch cultivation of E. coli, alternative cultivation/induction strategies remain to be more deeply exploited. Among these strategies, it could be mentioned the use of complex media with combination of different carbon sources, novel induction procedures and feed flow rate control matching the actual cell growth rate. Here, four different carbon source combinations (glucose, glycerol, glucose + glycerol and auto-induction) in batch media formulation were compared. A balanced combination of glucose and glycerol in a complex medium formulation led to: fast growth in the batch-phase; reduced plasmid instability by preventing early expression leakage; and protein volumetric productivity of 0.40 g.L-1.h-1. Alternative induction strategies were also investigated. A mixture of lactose and glycerol as supplementary medium fully induced a high biomass population, reaching a good balance between specific protein production (0.148 gprot.gDCW-1) and volumetric productivity (0.32 g.L-1.h-1). The auto-induction protocol showed excellent results on specific protein production (0.158 gprot.gDCW-1) in simple batch cultivations. An automated feed control based on the on-line estimated growth rate was implemented, which allowed cells to grow at higher rates than those generally used to avoid metabolic overflow, without leading to acetate accumulation. Some of the protocols described here may provide a useful alternative to standard cultivation and recombinant protein production processes, depending on the performance index that is expected to be optimized. The protocols using glycerol as carbon source and induction by lactose feeding, or glycerol plus glucose in batch medium and induction by lactose pulse led to rSpaA production in the range of 6 g.L-1, in short fed-batch processes (16 to 20 h) with low accumulation of undesired side metabolites.

show abstract

“…The gas valve actuator (SMC, model VY1100) and the pressure sensor (SMC, model ISE30A-01-F) were installed in the gas outlet line. The data were acquired using a Compact FieldPoint controller (National Instruments, model cFP2020) and SuperSys_HCDC software [19]. The dissolved oxygen tension was determined using an amperometric probe (Mettler Toledo, model InPro 6830) bearing a silicone membrane (model S96).…”

Section: Equipmentmentioning

confidence: 99%

Oxygen transfer in a pressurized airlift bioreactor

Campani

Ribeiro

Horta

et al. 2015

Bioprocess Biosyst Eng

View full text Add to dashboard Cite

Airlift bioreactors (ALBs) offer advantages over conventional systems, such as simplicity of construction, reduced risk of contamination, and efficient gas-liquid dispersion with low power consumption. ALBs are usually operated under atmospheric pressure. However, in bioprocesses with high oxygen demand, such as high cell density cultures, oxygen limitation may occur even when operating with high superficial gas velocity and air enriched with oxygen. One way of overcoming this drawback is to pressurize the reactor. In this configuration, it is important to assess the influence of bioreactor internal pressure on the gas hold-up, volumetric oxygen transfer coefficient (k(L)a), and volumetric oxygen transfer rate (OTR). Experiments were carried out in a concentric-tube airlift bioreactor with a 5 dm(3) working volume, equipped with a system for automatic monitoring and control of the pressure, temperature, and inlet gas flow rate. The results showed that, in disagreement with previous published results for bubble column and external loop airlift reactors, overpressure did not significantly affect k(L)a within the studied ranges of pressure (0.1-0.4 MPa) and superficial gas velocity in the riser (0.032-0.065 m s(-1)). Nevertheless, a positive effect on OTR was observed: it increased up to 5.4 times, surpassing by 2.3 times the oxygen transfer in a 4 dm(3) stirred tank reactor operated under standard cultivation conditions. These results contribute to the development of non-conventional reactors, especially pneumatic bioreactors operated using novel strategies for oxygen control.

show abstract

A supervision and control tool based on artificial intelligence for high cell density cultivations

Cited by 21 publications

References 28 publications

Pulmonary dry powder vaccine of pneumococcal antigen loaded nanoparticles

Pulmonary dry powder vaccine of pneumococcal antigen loaded nanoparticles

Non-conventional induction strategies for production of subunit swine erysipelas vaccine antigen in rE. coli fed-batch cultures

Oxygen transfer in a pressurized airlift bioreactor

Contact Info

Product

Resources

About