We have investigated the sensing performance of protein-based microcantilever biosensors prepared front multiple surface conjugation chemistries. The 11-mercaptoundecanoic acid monolayers were prepared according to both traditional and modified processes. In three protein-based biosensors, the modified process improved microcantilever-sensing performance by increasing the bending amplitude, a critical step toward developing a cost-effective microcantilever-based sensor platform for medical diagnostics and environmental and drug screening applications. Scanning electron microscopy (SEM) images demonstrated that proteins immobilized on the microcantilever Surface using the modified chemistry approach formed a compact layer.