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Towards background‐free serum analysis
Author(s) -
Danielsson Ake,
Markova Natalia,
Ohman Johan
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.lb156
Subject(s) - chromatography , chemistry , surface plasmon resonance , sample preparation , monoclonal antibody , antibody , materials science , nanotechnology , biology , immunology , nanoparticle
A fast and automatable sample preparation workflow for the determination of monoclonal antibodies (mAb's) in human blood serum samples using surface plasmon resonance, (SPR) was developed. Using standard sample preparation protocols, we frequently experienced high backgrounds in the analyses. The developed sample preparation workflow comprised two steps. The first step involved affinity capture of the target protein using magnetic beads with Protein G (Protein G Mag Sepharose™). The second step involved buffer exchange using a 96‐well filter plate, PD MultiTrap™G‐25, to a suitable buffer for analysis. To assess the stability of the target protein after sample preparation we used differential scanning calorimetry. The concentration of the target mAb, (Rituximab) in serum samples was determined using a Biacore™ T200 instrument. The results showed a reproducible reduction of the background signals in SPR analysis and an overall target protein recovery above 80%. The developed sample preparation workflow was successfully applied to determine Rituximab in human blood serum samples.