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In-line fiber optic...
In-line fiber optical sensor for detection of IgG aggregates in affinity chromatography
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- Tran, Thuy (författare)
- Linköpings universitet,Biofysik och bioteknik,Tekniska fakulteten,ArgusEye AB, Fridtunagatan 24, S-58213 Linkoping, Sweden,Laboratory of Molecular Materials
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- Gustavsson Lindberg, Robert (författare)
- Linköpings universitet,Biofysik och bioteknik,Tekniska fakulteten,Laboratory of Molecular Materials
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- Martinsson, Erik (författare)
- ArgusEye AB, Fridtunagatan 24, S-58213 Linkoping, Sweden
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- Bergqvist, Filip (författare)
- Cytiva Sweden AB, Sweden
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- Axen, Andreas (författare)
- Cytiva Sweden AB, Sweden
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- Lundström, Ingemar (författare)
- Linköpings universitet,Sensor- och aktuatorsystem,Tekniska fakulteten
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- Mandenius, Carl-Fredrik (författare)
- Linköpings universitet,Biofysik och bioteknik,Tekniska fakulteten,Laboratory of Molecular Materials
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- Aili, Daniel (författare)
- Linköpings universitet,Biofysik och bioteknik,Tekniska fakulteten,Laboratory of Molecular Materials
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(creator_code:org_t)
- ELSEVIER, 2024
- 2024
- Engelska.
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Ingår i: Journal of Chromatography A. - : ELSEVIER. - 0021-9673 .- 1873-3778. ; 1730
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Therapeutic monoclonal antibodies (mAbs) are critical for treatment of a wide range of diseases. Immunoglobulin G (IgG) is the most predominant form of mAb but is prone to aggregation during production. Detection and removal of IgG aggregates are time-consuming and laborious. Chromatography is central for purification of biopharmaceuticals in general and essential in the production of mAbs. Protein purification systems are usually equipped with detectors for monitoring pH, UV absorbance, and conductivity, to facilitate optimization and control of the purification process. However, specific in-line detection of the target products and contaminating species, such as aggregates, is currently not possible using convectional techniques. Here we show a novel fiber optical in-line sensor, based on localized surface plasmon resonance (LSPR), for specific detection of IgG and IgG aggregates during affinity chromatography. A flow cell with a Protein A sensor chip was connected to the outlet of the affinity column connected to three different chromatography systems operating at lab scale to pilot scale. Samples containing various IgG concentrations and aggregate contents were analyzed in-line during purification on a Protein A column using both pH gradient and isocratic elution. Because of avidity effects, IgG aggregates showed slower dissociation kinetics than monomers after binding to the sensor chips. Possibilities to detect aggregate concentrations below 1 % and difference in aggregate content smaller than 0.3 % between samples were demonstrated. In-line detection of aggregates can circumvent time-consuming off-line analysis and facilitate automation and process intensification.
Ämnesord
- NATURVETENSKAP -- Kemi -- Analytisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Analytical Chemistry (hsv//eng)
Nyckelord
- In -line sensor; Monoclonal antibodies; LSPR; Chromatography; IgG aggregate; Process analytical technology
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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