Multidrug resistant Pseudomonas aeruginosa is an alarming and emerging public health problem globally across the developing countries. Pseudomonas aeruginosa is still a major cause for nosocomial infection and approx 10-20% of these patients are admitted to the ICU’s. Bacterial isolates those are biofilm producers are more drug resistant than biofilm Non-producers. The aim of the present study was to evaluate the production of biofilm and β-lactamases (ESBL, MBL, AmpC) in multi drug resistant Pseudomonas aeruginosa isolated from ICU patients. The present cross-sectional prospective study was carried out in the Department of Microbiology, Santosh Medical College & Hospital, Ghaziabad, Uttar-Pradesh, India. A total of 115 isolates of P. aeruginosa were isolated from 502 clinical samples.  After confirmation of MDR status of P. aeruginosa further processing for biofilm and beta lactamases was performed accordingly. Biofilm production was done by test tube method and tissue culture plate method along with phenotypic profiling of ESBL, MBL and AmpC was performed by disc potentiation test; IMP-EDTA combined disc test and Cefoxitin Cloxacillin Double Disc synergy test (CC-DDST) respectively. Out of 502 total human clinical samples 115 isolates were Pseudomonas aeruginosa giving the prevalence rate of 23%. Among 115 isolates of P. aeruginosa 60 (52%) were MDR phenotypes, Out of 60 MDR isolates 23 (38.3%) were ESBL producers, 22 (36.6%) were MBL producers, and 3(5%) were AmpC producers. Out of total 115 isolates 68(59%) isolates were biofilm producers and 47 (40.8%) were biofilm non-producers. Strict antibiotic policies with early detection of beta lactamases and detection of biofilm production should be performed regularly for all clinical isolates of Pseudomonas aeruginosa so as to guide antibiotic selection along with better management of severe infection in ICU patients.