Direct analysis of mAb aggregates in mammalian cell culture supernatant

Paul, Albert Jesuran; Schwab, Karen; Hesse, Friedemann

doi:10.1186/s12896-014-0099-3

Cited by 41 publications

(44 citation statements)

References 40 publications

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“…Another explanation could be mAb aggregate precipitation during sample preparation. This was already seen by Paul et al (2014) [25], who described that large aggregates can be formed. These higher molecular weight species might be removed from the sample through centrifugation or filtration.…”

Section: Chemometric Modelingsupporting

confidence: 63%

“…The mAb aggregate concentration in the cell culture samples was determined with SE-HPLC using the Agilent 1100 system (Agilent Technologies, Santa Clara, CA, USA). A MAbPac SEC-1 column (Thermo Fisher Scientific, Waltham, MA, USA) was used, following the method described by Paul et al (2014) for the direct determination of mAb aggregate concentration in cell culture samples [25]. L-glutamine and glucose concentrations were determined enzymatically with the KonelabTM 20 XT (Thermo Fisher Scientific, Waltham, MA, USA) using the L-glutamine kit (Thermo Fisher Scientific, Waltham, MA, USA) and the Glucose HK kit (Thermo Fisher Scientific, Waltham, MA, USA), respectively.…”

Section: Offline Analyticsmentioning

confidence: 99%

“…Culture conditions as well as dye and substrate concentrations in medium and feed are stated in Table 1. Samples were drawn from the bioreactor, and product concentrations and aggregate levels were determined following the method described by Paul et al (2014) [25]. The PLSR models were calibrated with data recorded after dye addition.…”

Section: Online Monitoring Based On Extrinsic Fluorescencementioning

confidence: 99%

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Hybrid Modelling and Multi- Parametric Control of Bioprocesses

2018

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Section: Chemometric Modelingsupporting

confidence: 63%

Section: Offline Analyticsmentioning

confidence: 99%

Section: Online Monitoring Based On Extrinsic Fluorescencementioning

confidence: 99%

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Hybrid Modelling and Multi- Parametric Control of Bioprocesses

2018

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“…Cell concentration (Table 2) and soluble aggregate formation (Table 3) remained unchanged, whereas in combination with osmolality a synergistic positive effect on specific mAb productivity (Table 2) was observed. It is known that the aggregation rate of proteins is strongly influenced by the pH (Arosio, Rima, & Morbidelli, 2013;Chi, Krishnan, Randolph, & Carpenter, 2003;Paul et al, 2014). It is known that the aggregation rate of proteins is strongly influenced by the pH (Arosio, Rima, & Morbidelli, 2013;Chi, Krishnan, Randolph, & Carpenter, 2003;Paul et al, 2014).…”

Section: Initial Ph Showed Only Marginal Effectsmentioning

confidence: 99%

Identification of process conditions influencing protein aggregation in Chinese hamster ovary cell culture

Paul

Handrick

Ebert

et al. 2018

Biotech & Bioengineering

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Protein aggregation of monoclonal antibodies (mAbs) is a common phenomenon associated with the production of these biopharmaceuticals. These aggregates can lead to adverse side effects in patients upon administration, thus expensive downstream processing steps to remove the higher molecular weight species are inevitable. A preferable approach is to reduce the level of aggregation during bioprocessing by a careful adjustment of critical process parameters. Recently, new analytical methods enabled characterization of mAb aggregation during bioprocessing of mammalian cells. Furthermore, rapid and efficient bioprocess optimization has been performed using design of experiments (DoE) strategies. In this work, we describe a DoE-based approach for the analysis of process parameters and cell culture additives influencing protein aggregation in Chinese hamster ovary (CHO) cell cultures. Important bioprocess variables influencing the aggregation of mAb and host cell proteins were identified in initial screening experiments. Response surface modeling was further applied in order to find optimal conditions for the reduction of protein aggregation during cell culture. It turned out that a temperature-shift to 31 °C, osmolality above 420 mOsm/kg, agitation at 100 rpm and 0.04% (w/v) antifoam significantly reduced the level of aggregates without substantial detrimental effects on cell culture performance in our model system. Finally, the aggregation reducing conditions were verified and applied to another production system using a different bioprocess medium and another CHO cell line producing another mAb. Our results show that protein aggregation can be controlled during cell culture and helps to improve bioprocessing of mAbs, by giving insights into the protein aggregation at its origin in mammalian cell culture.

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“…Purification processes are in general capable to sufficiently reduce the aggregation level of the cell culture broth 169 .…”

Section: Aggregatesmentioning

confidence: 99%

Tailoring recombinant protein quality by rational media design

Brühlmann

Jordan

Hemberger³

et al. 2015

Biotechnology Progress

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Clinical efficacy and safety of recombinant proteins are closely associated with their structural characteristics. The major quality attributes comprise glycosylation, charge variants (oxidation, deamidation, and C‐ & N‐terminal modifications), aggregates, low‐molecular‐weight species (LMW), and misincorporation of amino acids in the protein backbone. Cell culture media design has a great potential to modulate these quality attributes due to the vital role of medium in mammalian cell culture. The purpose of this review is to provide an overview of the way both classical cell culture medium components and novel supplements affect the quality attributes of recombinant therapeutic proteins expressed in mammalian hosts, allowing rational and high‐throughput optimization of mammalian cell culture media. A selection of specific and/or potent inhibitors and activators of oligosaccharide processing as well as components affecting multiple quality attributes are presented. Extensive research efforts in this field show the feasibility of quality engineering through media design, allowing to significantly modulate the protein function. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:615–629, 2015

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Direct analysis of mAb aggregates in mammalian cell culture supernatant

Cited by 41 publications

References 40 publications

Hybrid Modelling and Multi- Parametric Control of Bioprocesses

Hybrid Modelling and Multi- Parametric Control of Bioprocesses

Identification of process conditions influencing protein aggregation in Chinese hamster ovary cell culture

Tailoring recombinant protein quality by rational media design

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