Abstract: The molecular docking technology were used to investigate the combining ability of heat shock protein 27(HSP27)and β adrenergic receptor agonists and the possibility that heat shock protein 27 be used to detect β adrenergic receptor agonists. Artificially expressed heat shock protein 27 and β₂ adrenergic receptor(β₂AR)were combined with β adrenergic receptor agonists respectively by molecular docking technology. Based on the results of molecular docking,the differences of their combining ability were compared,and then verified by enzyme-linked immunosorbent assay(ELISA). The results showed that nine total score of heat shock protein 27 was higher than β₂ adrenergic receptor with fifteen small molecule drugs of β adrenergic receptor agonist in molecular docking. Enzyme-linked immunosorbent assay results showed that heat shock protein 27 could be reacted with all three horse radish peroxidase(HRP)-β adrenergic receptor agonists of clenbuterol,fenoterol,ractopamine specifically and the optical density(OD)values obtained were 0.685,0525,and 0.662 respectively. The results of enzyme-linked immunosorbent assay are in accordance with the results of molecular docking basically. In this paper,heat shock protein 27 has strong combining ability with β adrenergic agonists,which can be used to detect β adrenergic receptor agonists in all probability.