Epothilone B is a highly promising prospective anticancer agent. At present, the cost of producing epothilone B is very high mainly due to two reasons. The first the toxicity and feedback inhibition of epothilones lead to difficult to improve fermentation yield. The second is the fermentation broth contains a large number of epothilone homologues, leading to separation and purification costs of epothilone B are very high. Therefore, the development of epothilone B adsorption used in its fermentation and extraction will significantly reduce production costs. In the present study, Molecularly imprinted polymer(MIPs) of epothilone B was prepared by precipitation polymerization using three members of epothilone (epothilone A, epothilone B and epothilone D)as templates，methacrylic acid as functional monomer, and ethylene glycol dimethacrylate as cross-linker. MIPs can specifically recognise epothione B. By measuring the adsorbance of different mixed-templates and different ratio of templates/MAA, the best condition was determined. The results showed that the imprinting effect was best when the molar ratio of templates/MAA is 1:6 and mixed-templates are epo B and epo D (1:1), respectively. MIPs exhibited a high recognizable capacity to Epothilone B. Therefore, that was expected to be used as extraction and purification of Epothilone B in the process of fermentation.