The major hallmarks of Alzheimer's disease (AD) are the extracellular accumulation of pathological amyloid beta (Aβ) in the brain parenchyma and Aβ deposition in cerebral blood walls (cerebral amyloid angiopathy; CAA). Although CAA occurs in more than 80% of AD patients, the mechanisms of Aβ deposition and clearance around the vessel walls are unknown. We found Aβ-degrading activity in human serum during analysis of the regulatory mechanism of Aβ production in human endothelial cells. To elucidate the metabolic dynamics of Aβ surrounding the brain microvessels, we identified Aβ-degrading activity in human serum (blood Aβ-degrading activity: BADA) by column chromatography and LC/MS. BADA exhibited characteristics of an acidic protein, pI 4.3, which had two different protein surface charges (low and high affinity cations). Both BADA fractions had a relative molecular mass of greater than 400 kDa. Furthermore, BADA in the low affinity cation fraction was inhibited by the serine protease inhibitor 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF). We clarified alpha-2-macroglobulin (a2M) and several serine proteases from this BADA by LC-MS. Moreover, we demonstrated that BADA is increased by approximately 5000-fold in human serum by column chromatography. Therefore, BADA may play an important role in the circulation and metabolism of Aβ in human brain microvessels.