Determination of Recombinant Protein in Animal Cell Culture Supernatant by Near-Infrared Spectroscopy

Harthun, Sven; Matischak, Kathrin; Friedl, Peter

doi:10.1006/abio.1997.2218

Cited by 31 publications

(23 citation statements)

References 12 publications

(5 reference statements)

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“…Also, once the spectra had been deconvoluted (using the second derivative) the analyte signals are more dominant compared to the effect of temperature change, so it is still possible to determine analyte information from the spectra. This is significant from an industrial viewpoint because during many industrial processes the temperature is often increased or decreased in order to induce product pro- Once method of data collection selected not possible to change without disrupting the sterile environment of the bioreactor Usually degassed/temperature equilibrated samples Gas phase effects (bubbles) (Vaidyanathan et al, 1999a;Ge et al, 1994)/temperature change (Harthun et al, 1997) in some processes No fouling Probe fouling or blinding by adherent film Macaloney, 1996a) …”

Section: Transfer From At-line To On-linementioning

confidence: 98%

“…Temperature is known to have an effect on the spectra (Harthun et al, 1997). In this system there was a temperature increase during the course of a fermentation run.…”

Section: Transfer From At-line To On-linementioning

confidence: 99%

“…These conditions are much closer to actual industrial practice than those reported in previous on-line NIR studies. Since both these factors can have profound effects on matrix character (Ge et al, 1994;Harthun et al, 1997), overall application of NIRS in such a process is both highly challenging and more reflective of the real difficulties NIRS would face in being applied to industrial production processes.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Use of at‐line and in‐situ near‐infrared spectroscopy to monitor biomass in an industrial fed‐batch Escherichia coli process

Arnold

Gaensakoo²,

Harvey

et al. 2002

Biotech & Bioengineering

124

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One of the key goals in bioprocess monitoring is to achieve real-time knowledge of conditions within the bioreactor, i.e., in-situ. Near-infrared spectroscopy (NIRS), with its ability to carry out multi-analyte quantification rapidly with little sample presentation, is potentially applicable in this role. In the present study, the application of NIRS to a complex, fed-batch industrial E. coli (RV308/PHKY531) process was investigated. This process undergoes a series of temperature changes and is vigorously agitated and aerated. These conditions can pose added challenges to in-situ NIRS. Using the measurement of a key analyte (biomass) as an illustration, the details of the relationship between the at-line and in-situ use of NIRS are considered from the viewpoint of both theory and practical application. This study shows that NIRS can be used both at-line and in-situ in order to achieve good predictive models for biomass. There are particular challenges imposed by in-situ operation (loss of wavelength regions and noise) which meant the need for signal optimisation studies. This showed that whilst the at-line modelling process may provide some useful information for the in-situ process, there were distinct differences. This study shows that the in-situ use of NIRS in a highly challenging matrix (similar to those encountered in current industrial practice) is possible, and thus extends previous works in the area.

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Section: Transfer From At-line To On-linementioning

confidence: 98%

“…Temperature is known to have an effect on the spectra (Harthun et al, 1997). In this system there was a temperature increase during the course of a fermentation run.…”

Section: Transfer From At-line To On-linementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Use of at‐line and in‐situ near‐infrared spectroscopy to monitor biomass in an industrial fed‐batch Escherichia coli process

Arnold

Gaensakoo²,

Harvey

et al. 2002

Biotech & Bioengineering

124

View full text Add to dashboard Cite

show abstract

“…To date, in the field of biological processing NIRS has been mainly used for bacterial [9][10][11][12], microalgae [13] and fungal [14] cultures. In the case of animal cell culture processes, NIRS is currently not routinely used, with only few reports describing NIRS analysis of soluble components in cell culture supernatants [15][16][17][18]. All these studies were performed with CHO and insect cells cultivated in suspension mode, and similar work has not been reported for adherent cells.…”

Section: Introductionmentioning

confidence: 94%

In situ quantification of microcarrier animal cell cultures using near-infrared spectroscopy

Petiot

Bernard-Moulin²,

Magadoux

et al. 2010

Process Biochemistry

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“…For example, there are multiple biomanufacturing cases of the successful application of in-line sensors along with modeling methodology to monitor and control the growth of cell culture processes (Gnoth et al, 2007;Junker and Wang, 2006;Velut et al, 2007). Likewise, Near-Infrared spectroscopy has been applied to quantifying protein expression (Harthun et al, 1997) or RNA levels (Yeung et al, 1999) in cell cultures with good correlation to offline measurements, while DNA microarray tools have been evaluated for process quality applications in vaccine production (Streefland et al, 2007). Downstream chromatography operations have benefited from using on-line HPLC based applications to provide real-time pooling criteria that is difficult to automate based on the effluent absorbance (Fahrner et al, 1998;Rathore et al, 2008) as well as off-line HPLC to determine loading density (Fahrner and Blank, 1999).…”

Section: Introductionmentioning

confidence: 99%

Biomanufacturing process analytical technology (PAT) application for downstream processing: Using dissolved oxygen as an indicator of product quality for a protein refolding reaction

Pizarro¹,

Dinges²,

Sánchez³

et al. 2009

Biotech & Bioengineering

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Process analytical technology (PAT) is an initiative from the US FDA combining analytical and statistical tools to improve manufacturing operations and ensure regulatory compliance. This work describes the use of a continuous monitoring system for a protein refolding reaction to provide consistency in product quality and process performance across batches. A small-scale bioreactor (3 L) is used to understand the impact of aeration for refolding recombinant human vascular endothelial growth factor (rhVEGF) in a reducing environment. A reverse-phase HPLC assay is used to assess product quality. The goal in understanding the oxygen needs of the reaction and its impact to quality, is to make a product that is efficiently refolded to its native and active form with minimum oxidative degradation from batch to batch. Because this refolding process is heavily dependent on oxygen, the % dissolved oxygen (DO) profile is explored as a PAT tool to regulate process performance at commercial manufacturing scale. A dynamic gassing out approach using constant mass transfer (k(L)a) is used for scale-up of the aeration parameters to manufacturing scale tanks (2,000 L, 15,000 L). The resulting DO profiles of the refolding reaction show similar trends across scales and these are analyzed using rpHPLC. The desired product quality attributes are then achieved through alternating air and nitrogen sparging triggered by changes in the monitored DO profile. This approach mitigates the impact of differences in equipment or feedstock components between runs, and is directly inline with the key goal of PAT to "actively manage process variability using a knowledge-based approach."

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Determination of Recombinant Protein in Animal Cell Culture Supernatant by Near-Infrared Spectroscopy

Cited by 31 publications

References 12 publications

Use of at‐line and in‐situ near‐infrared spectroscopy to monitor biomass in an industrial fed‐batch Escherichia coli process

Use of at‐line and in‐situ near‐infrared spectroscopy to monitor biomass in an industrial fed‐batch Escherichia coli process

In situ quantification of microcarrier animal cell cultures using near-infrared spectroscopy

Biomanufacturing process analytical technology (PAT) application for downstream processing: Using dissolved oxygen as an indicator of product quality for a protein refolding reaction

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