title: Whole cell-based surface plasmon resonance measurement to assess binding of anti-TNF agents to transmembrane target.
abstract: We developed a technique for the measurement of surface plasmon resonance (SPR) to detect interactions of anti-tumor necrosis factor (TNF) agents with transmembrane TNF-α (mTNF-α) on living whole cells. The injection of a suspension of mTNF-α expressing Jurkat cells, used as an analyte, gave a clear binding response to anti-TNF agents, such as etanercept, infliximab and adalimumab, immobilized on sensorchip. The binding response of the analyte cells increased in a concentration-dependent manner and was competitively reduced by adding soluble TNF receptors to the analyte cell suspension. Treatment of analyte cells with free anti-TNF agent before injection reduced the binding response between the analyte cells and immobilized-etanercept on sensorchip, and the inhibitory effect of free anti-TNF agent was concordant with the affinity of anti-TNF agent for soluble TNF-α. These findings indicate that the SPR response arises from specific binding between anti-TNF agent and its target on cell membrane.
authors.affiliation: Biological Research Department, Sawai Pharmaceutical Co., Ltd., Osaka 532-0003, Japan. Electronic address: t.ogura@sawai.co.jp.
authors.forename: Takeharu
authors.initials: T
authors.lastname: Ogura
authors.affiliation: Biological Research Department, Sawai Pharmaceutical Co., Ltd., Osaka 532-0003, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Tanaka
authors.affiliation: Biological Research Department, Sawai Pharmaceutical Co., Ltd., Osaka 532-0003, Japan.
authors.forename: Hiromu
authors.initials: H
authors.lastname: Toyoda
mesh_terms: D000068879:Adalimumab / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism; D018501:Antirheumatic Agents / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D000068800:Etanercept / Q000378:metabolism; D006801:Humans; D000069285:Infliximab / Q000378:metabolism; D019169:Jurkat Cells; D011485:Protein Binding; D020349:Surface Plasmon Resonance; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D018501:Antirheumatic Agents; D002352:Carrier Proteins; D014409:Tumor Necrosis Factor-alpha; D000069285:Infliximab; D000068879:Adalimumab; D000068800:Etanercept
keywords: Anti-TNF agent; Biological cell–protein interaction; Surface plasmon resonance; Transmembrane TNF-α