Wound biofilms pose a great clinical challenge. Herein, we report a dissolvable microneedle patch for dual delivery of monoclonal antibodies anti‐PBP2a and engineered antimicrobial peptides W379. in vitro antibacterial efficacy testing with microneedle patches containing a combination of 250 ng/mL W379 and 250 ng/mL anti‐BPB2a decreased the bacterial count from ∼3.31 × 107 CFU/mL to 1.28 × 102 CFU/mL within 2 h without eliciting evident cytotoxicity. ex vivo testing indicates W379 and anti‐PBP2a co‐loaded microneedle patch displayed a remarkable reduction of bacterial load by approximately 7.18 log CFU after administered only once within 48 h. The bacterial count was significantly diminished compared to the treatment by either W379 or anti‐PBP2a‐loaded alone microneedle patches. When administered twice within 48 h, no bacteria were identified. Further in vivo study also revealed that after two treatments of W379 and anti‐PBP2a co‐loaded PVP microneedle patches within 48 h, the bacterial colonies were undetectable in a type II diabetic mouse wound biofilm model. Taken together, W379 and anti‐PBP2a co‐loaded PVP microneedle patches hold great promise in treating wound biofilms.This article is protected by copyright. All rights reserved