title: Nine surface plasmon resonance assays for specific protein quantitation during cell culture and process development.
abstract: 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 13 days 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 12 days, allowing specific protein concentration measurement of a sample to be completed at line within 10 min. 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.
authors.affiliation: GE Healthcare BioSciences, SE-751 84 Uppsala, Sweden. Electronic address: asa.frostell@ge.com.
authors.forename: Åsa
authors.initials: Å
authors.lastname: Frostell
authors.affiliation: GE Healthcare BioSciences, SE-751 84 Uppsala, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Mattsson
authors.affiliation: GE Healthcare BioSciences, SE-751 84 Uppsala, Sweden.
authors.forename: Åsa
authors.initials: Å
authors.lastname: Eriksson
authors.affiliation: GE Healthcare BioSciences, SE-751 84 Uppsala, Sweden.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Wallby
authors.affiliation: GE Healthcare BioSciences, SE-751 84 Uppsala, Sweden.
authors.forename: Johan
authors.initials: J
authors.lastname: Kärnhall
authors.affiliation: GE Healthcare BioSciences, SE-751 84 Uppsala, Sweden.
authors.forename: Nina B
authors.initials: NB
authors.lastname: Illarionova
authors.affiliation: GE Healthcare BioSciences, SE-751 84 Uppsala, Sweden.
authors.forename: Camilla
authors.initials: C
authors.lastname: Estmer Nilsson
mesh_terms: D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000032:analysis; D001798:Blood Proteins / Q000032:analysis* / Q000276:immunology; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D001798:Blood Proteins
keywords: Albumin; IgA; IgG; Quantitate; SPR; Transferrin