The present disclosure provides an anisotropic conductive film, a display panel, and manufacturing method thereof. The anisotropic conductive film includes a conductive particle, the conductive particle including a covalent organic framework material, the covalent organic framework material including PyVg-COF. The display panel including a first substrate, a second substrate and an anisotropic conductive film. The method of manufacturing the display panel including: providing a first substrate, coating an anisotropic conductive film on the first substrate, coupling the second substrate to the first substrate, and bonding the first substrate to the second substrate. The present disclosure provides the conductive particles of the covalent organic frame material replace the existing gold ball as an anisotropic conductive film for bonding, thereby saving the manufacturing cost of the gold ball, improving the conductivity and water resistance, and avoiding the bonding contact point being oxidation.