Recent developments in mass spectrometry-based targeted proteomics for clinical cancer biomarkers have shown significant potential for improving cancer diagnosis, monitoring, and treatment. Targeted proteomics, particularly using liquid chromatography-tandem mass spectrometry (LC-MS/MS), offers high specificity and sensitivity for quantifying cancer biomarkers in biological samples, such as blood and urine. This technique allows for the precise measurement of proteins, which are critical for understanding cancer pathophysiology. Unlike immunoassays, which can be affected by interferences, LC-MS/MS provides more reliable results by directly measuring target proteins and their fragments. Targeted proteomics has several advantages, including high reproducibility, the ability to quantify multiple biomarkers simultaneously, and the capacity to handle complex biological matrices. However, challenges remain, such as the need for high technical proficiency, high costs, and the complexity of the workflows involved. Recent advancements in automation, software capabilities, and sample preparation techniques have improved the efficiency and accessibility of targeted proteomics. For example, automated liquid handling systems and improved software for data analysis have enhanced the throughput and accuracy of LC-MS/MS assays. Despite these improvements, the clinical implementation of targeted proteomics is still limited due to the lack of standardized validation protocols and the need for robust clinical validation studies. The development of targeted proteomics assays requires careful method development, validation, and implementation to meet regulatory requirements. Additionally, the integration of targeted proteomics into clinical laboratories requires coordination between various departments, including laboratory management, quality control, and information technology. Future directions for targeted proteomics include the development of more automated systems, the use of immunoaffinity enrichment to improve sensitivity, and the application of multidimensional LC and ion mobility spectrometry to enhance the resolution and sensitivity of proteomic analyses. These advancements are expected to increase the utility of targeted proteomics in clinical settings, enabling more accurate and efficient cancer biomarker analysis. Overall, targeted proteomics holds great promise for improving cancer diagnostics and personalized treatment strategies.Recent developments in mass spectrometry-based targeted proteomics for clinical cancer biomarkers have shown significant potential for improving cancer diagnosis, monitoring, and treatment. Targeted proteomics, particularly using liquid chromatography-tandem mass spectrometry (LC-MS/MS), offers high specificity and sensitivity for quantifying cancer biomarkers in biological samples, such as blood and urine. This technique allows for the precise measurement of proteins, which are critical for understanding cancer pathophysiology. Unlike immunoassays, which can be affected by interferences, LC-MS/MS provides more reliable results by directly measuring target proteins and their fragments. Targeted proteomics has several advantages, including high reproducibility, the ability to quantify multiple biomarkers simultaneously, and the capacity to handle complex biological matrices. However, challenges remain, such as the need for high technical proficiency, high costs, and the complexity of the workflows involved. Recent advancements in automation, software capabilities, and sample preparation techniques have improved the efficiency and accessibility of targeted proteomics. For example, automated liquid handling systems and improved software for data analysis have enhanced the throughput and accuracy of LC-MS/MS assays. Despite these improvements, the clinical implementation of targeted proteomics is still limited due to the lack of standardized validation protocols and the need for robust clinical validation studies. The development of targeted proteomics assays requires careful method development, validation, and implementation to meet regulatory requirements. Additionally, the integration of targeted proteomics into clinical laboratories requires coordination between various departments, including laboratory management, quality control, and information technology. Future directions for targeted proteomics include the development of more automated systems, the use of immunoaffinity enrichment to improve sensitivity, and the application of multidimensional LC and ion mobility spectrometry to enhance the resolution and sensitivity of proteomic analyses. These advancements are expected to increase the utility of targeted proteomics in clinical settings, enabling more accurate and efficient cancer biomarker analysis. Overall, targeted proteomics holds great promise for improving cancer diagnostics and personalized treatment strategies.