Automated process monitoring of monoclonal antibody production

Paliwal, Sandeep K.; Nadler, Timothy; Wang, Daniel I. C.; Regnier, Fred E.

doi:10.1021/ac00071a005

Cited by 33 publications

(14 citation statements)

References 5 publications

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“…Chromatographic assays are fast [27] and can be used to quickly assay recombinant proteins [28], and commercial instrumentation speci®-cally designed to run on-line chromatographic assays is available [29]. Perfusion chromatography can produce protein analysis in as little as 20 seconds [30,31] and protein A is antibody-speci®c, so the on-line assay used protein A immobilized on perfusion chromatography media, providing a rapid antibody-speci®c assay [32]. Using an on-line assay makes it possible to gather the necessary empirical data much faster than would be possible otherwise.…”

Section: Analytical Chromatographymentioning

confidence: 99%

The optimal flow rate and column length for maximum production rate of protein A affinity chromatography

Fahrner¹,

Iyer²,

Blank³

1999

Bioprocess Engineering

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Production rate is an important parameter in the design of ef®cient protein A af®nity chromatography processes for purifying recombinant monoclonal antibodies. A simple equation was derived that expresses production rate in terms of¯ow rate and column length. Changes in¯ow rate and column length will not affect the antibody and are therefore easily varied for bioprocess applications. In the equation, production rate depends on dynamic capacity, which can be expressed as a function of the load¯ow rate and column length. The only empirical data needed for production rate optimization is the relationship of dynamic capacity to load¯ow rate and column length, which was quickly determined by using an on-line assay. The optimal production rate was found at a high ow rate, a low column length, and a low dynamic capacity, which has several implications for using high production rate protein A af®nity chromatography for antibody manufacturing.

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Section: Analytical Chromatographymentioning

confidence: 99%

The optimal flow rate and column length for maximum production rate of protein A affinity chromatography

Fahrner¹,

Iyer²,

Blank³

1999

Bioprocess Engineering

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“…In order to minimize human impact, automated sampling can be applied. Automated analytical chromatography has been used in upstream processing to monitor the mAb titers (Chase, ; Ozturk et al, ; Paliwal et al, ). In downstream processing, this technique was successfully used for mAb quantification in the column effluent during the load phase of Protein A chromatography.…”

Section: Introductionmentioning

confidence: 99%

Real‐time monitoring and control of the load phase of a protein A capture step

Rüdt

Brestrich

Rolinger

et al. 2016

Biotech & Bioengineering

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The load phase in preparative Protein A capture steps is commonly not controlled in real‐time. The load volume is generally based on an offline quantification of the monoclonal antibody (mAb) prior to loading and on a conservative column capacity determined by resin‐life time studies. While this results in a reduced productivity in batch mode, the bottleneck of suitable real‐time analytics has to be overcome in order to enable continuous mAb purification. In this study, Partial Least Squares Regression (PLS) modeling on UV/Vis absorption spectra was applied to quantify mAb in the effluent of a Protein A capture step during the load phase. A PLS model based on several breakthrough curves with variable mAb titers in the HCCF was successfully calibrated. The PLS model predicted the mAb concentrations in the effluent of a validation experiment with a root mean square error (RMSE) of 0.06 mg/mL. The information was applied to automatically terminate the load phase, when a product breakthrough of 1.5 mg/mL was reached. In a second part of the study, the sensitivity of the method was further increased by only considering small mAb concentrations in the calibration and by subtracting an impurity background signal. The resulting PLS model exhibited a RMSE of prediction of 0.01 mg/mL and was successfully applied to terminate the load phase, when a product breakthrough of 0.15 mg/mL was achieved. The proposed method has hence potential for the real‐time monitoring and control of capture steps at large scale production. This might enhance the resin capacity utilization, eliminate time‐consuming offline analytics, and contribute to the realization of continuous processing. Biotechnol. Bioeng. 2017;114: 368–373. © 2016 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals, Inc.

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“…Protein A affinity chromatography has been used as a fast analytical tool in the quantification of monoclonal antibodies (mAb) . Because mAb and protein A have high affinity for each other, acidic pH conditions are required to elute mAb from protein A affinity columns.…”

Section: Introductionmentioning

confidence: 99%

“…Protein A affinity chromatography has been used as a fast analytical tool in the quantification of monoclonal antibodies (mAb). 1,2 Because mAb and protein A have high affinity for each other, acidic pH conditions are required to elute mAb from protein A affinity columns. Protein A affinity chromatography can also be used by combining with size-exclusion chromatography as a single high throughput analytical tool for simultaneous quantification of mAb, mAb aggregate, and host cell proteins in cell culture supernatants.…”

Section: Introductionmentioning

confidence: 99%

Arginine as an eluent overcomes the hindrance of monoclonal antibody quantification by dextran sulfate in protein A affinity chromatography

Kim

Park

2015

Biotechnology Progress

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Analytical chromatography using protein A affinity columns was employed for the fast and simple quantitative analysis of monoclonal antibodies (mAb) from suspension cultures of recombinant Chinese hamster ovary (rCHO) cells. Reliable results could not be obtained from analysis of rCHO cell culture supernatants containing dextran sulfate using elution buffers such as phosphate, glycine, or MgCl2. These problems increased as the number of analysis and the concentration of dextran sulfate in samples increased. Arginine was identified as an alternative eluent to overcome the hindrance by dextran sulfate. When the samples contain dextran sulfate up to 100 mg/L, the elution buffer containing 0.6–1.0 M arginine at pH 3.0–3.8 is useful for the effective analysis. Reproducible results in the mAb quantification could be obtained by this developed arginine elution buffer from rCHO cell culture supernatants containing dextran sulfate. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1536–1541, 2015

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Automated process monitoring of monoclonal antibody production

Cited by 33 publications

References 5 publications

The optimal flow rate and column length for maximum production rate of protein A affinity chromatography

The optimal flow rate and column length for maximum production rate of protein A affinity chromatography

Real‐time monitoring and control of the load phase of a protein A capture step

Arginine as an eluent overcomes the hindrance of monoclonal antibody quantification by dextran sulfate in protein A affinity chromatography

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