Nine surface plasmon resonance assays for specific protein quantitation during cell culture and process development

Quantitation of protein is essential during pharmaceutical development, and a variety of methods and technologies for determination of total and specific protein concentration are available. Here we describe the development of a streamlined assay platform for specific quantitation assays using surfa...

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Bibliographic Details
Published in:Analytical biochemistry 2015-05, Vol.477, p.1-9
Main Authors: Frostell, Åsa, Mattsson, Anna, Eriksson, Åsa, Wallby, Elisabeth, Kärnhall, Johan, Illarionova, Nina B., Estmer Nilsson, Camilla
Format: Article
Language:English
Subjects:
Albumin
Animals
Antibodies, Immobilized - chemistry
Antibodies, Immobilized - immunology
Antibodies, Monoclonal - analysis
Blood Proteins - analysis
Blood Proteins - immunology
CHO Cells
Cricetinae
Cricetulus
Humans
IgA
IgG
Quantitate
SPR
Surface Plasmon Resonance - methods
Transferrin
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Summary:Quantitation of protein is essential during pharmaceutical development, and a variety of methods and technologies for determination of total and specific protein concentration are available. Here we describe the development of a streamlined assay platform for specific quantitation assays using surface plasmon resonance (SPR) technology. A total of nine different assays were developed using similar conditions, of which eight assays were for quantitation of different human blood plasma proteins (IgG, IgG1–4 subclasses, IgA, transferrin, and albumin) from a chromatography-based IgG plasma process. Lastly, an assay for monitoring the concentration of a recombinant monoclonal antibody during 13days of CHO cell culturing was developed. Assay performances were compared with enzyme-linked immunosorbent assay (ELISA), nephelometry, ARCHITECT, and Cobas c501. SPR assays were shown to have higher sensitivity than analysis using nephelometry, ARCHITECT, and Cobas and to have significantly lower analysis and hands-on time compared with ELISA. Furthermore, the SPR assays were robust enough to be used for up to 12days, allowing specific protein concentration measurement of a sample to be completed at line within 10min. Using the same platform with only few varied parameters between different assays has saved time in the lab as well as for evaluation and presentation of results.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2015.02.010