Screening and quantification of TNF-α ligand from Angelicae Pubescentis Radix by biosensor and UPLC-MS/MS

The aim of this study is to establish a method for rapid screening of active ingredients targeting TNF-alpha from Chinese herbal medicines. Take Angelicae Pubescentis Radix (APR) as an example, surface plasma resonance technique was used to establish for screening small molecule inhibitors of TNF-alpha from APR extract. Then UPLC-MS/MS coupled with chemometric was used for quantitative and evaluate the differences of the candidate compounds bound to TNF-alpha in APR from different sources. In the experiment, TNF-alpha protein was fixed on the CM5 chip surface of biacore T200 biosensor by amino coupling. A series of small molecular compounds in APR were screened and six phenolic acid compounds had a strong affinity for TNF-alpha protein and could be used as TNF-alpha antagonists. In summary, the targeted drug screening method for TNF-alpha protein based on SPR technology established in this study can be used to screen anti-TNF-alpha small molecule inhibitors. UPLC-MS/MS can accurately quantify 15 active ingredients, which provides reliable experimental data and new research ideas for targeted drug research on TNF-alpha protein.