Abstract <p>The most prevalent protein in the bloodstream, serum albumins—mainly human serum albumin (HSA) and bovine serum albumin (BSA)—are essential to the pharmacokinetics of numerous medications, including antibiotics. HSA is a crucial modulator of drug transport, distribution, and efficacy due to its broad range of endogenous and exogenous chemical binding capabilities. Antibiotic interactions with serum albumins are particularly significant as they directly influence the bioavailability of the free, pharmacologically active form of the drug. High binding affinity to albumin often results in prolonged half-life but reduced therapeutic action due to limited free drug concentration, whereas low binding may lead to increased clearance and potential toxicity. Understanding these interactions is essential for precise dose adjustment, especially in clinical scenarios involving altered albumin levels such as liver dysfunction, renal disease, inflammation, or critical illness. Various spectroscopic techniques—such as fluorescence quenching, circular dichroism, and UV-visible absorption spectroscopy—alongside molecular docking and simulation studies have been instrumental in characterizing the binding mechanisms, sites, and conformational changes induced in albumin upon interaction with antibiotics. Moreover, differences in binding affinity based on the class of antibiotic (e.g., β-lactams, aminoglycosides, fluoroquinolones, lincosamide) and structural properties further complicate dosing regimens. Advances in computational chemistry and personalized medicine offer promising tools to predict and optimize these interactions for individual patients. This review emphasizes the biochemical, biophysical, and clinical relevance of serum albumin–antibiotic interactions and highlights the need for continued interdisciplinary research to refine pharmacological models and improve therapeutic outcomes.</p>