Under the present investigation, effect of ciprofloxacin (CIP) on Escherichia coli has been investigated using electrochemical, spectroscopic and atomic force microscope (AFM) measurements. Investigation reveals the interaction pattern of CIP with E. coli. The CIP essentially interacts with the outer membrane protein F (OmpF), the formation constant of the complex forms between CIP and the OmpF active sites over E. coli is obtained as log Kf of 12.1. Spectroscopic measurements are carried out, which supports the electrochemical measurements on the interaction between CIP and E. Coli , at a higher concentration, CIP induces lysis of the E. coli cell membrane. Spectroscopic investigations further reveals that the Fee S containing proteins present inside the E. coli cells released out through the ruptured cell membrane of E. coli. Different degrees of detrimental effects on E. coli has been observed when exposed to different concentrations of the drugs. The microscopic images obtained from the AFM scans of E. coli in presence of CIP shows deformation of the E. coli cell wall and its rupture with increasing concentrations of CIP .