Abstract
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a robust analytical technology that has become integral to biomolecular research. Since its introduction into microbiology in the early 2000s, its versatility has enabled a wide spectrum of applications extending from routine microbial identification to advanced proteomic profiling, antimicrobial resistance testing, biomarker discovery, and even historical disease investigation. In proteomics, MALDI-TOF MS has proven valuable for identifying disease-associated proteins, with applications in oncology, metabolic disorders such as diabetes and dyslipidemia, neurodegenerative diseases, hemoglobinopathies, and neonatal screening. Additionally, it has facilitated pharmacokinetic studies by enabling detailed analysis of drug distribution and metabolism. Despite limitations such as dependency on reference databases and challenges in distinguishing closely related species, ongoing advancements continue to enhance its accuracy and range. The integration of MALDI-TOF MS with molecular methods like polymerase chain reaction further strengthens its diagnostic utility. This review aims to present recent technological progress while highlighting the expanding interdisciplinary utility of MALDI-TOF MS. Emphasis is placed on emerging fields, including paleopathology, where its potential remains underexploited. By outlining its evolving capabilities, we propose a conceptual framework that positions MALDI-TOF MS as a unifying platform capable of driving innovation across diverse scientific and biomedical disciplines.