Whole cell-based surface plasmon resonance measurement to assess binding of anti-TNF agents to transmembrane target

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-alpha (mTNF-alpha) on living whole cells. The injection of a suspension of mTNF-alpha 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-alpha. These findings indicate that the SPR response arises from specific binding between anti-TNF agent and its target on cell membrane. (C) 2016 Elsevier Inc. All rights reserved.