A novel detection method of cleaved plasma high‐molecular‐weight kininogen reveals its correlation with Alzheimer's pathology and cognitive impairment

Abstract

Abstract Introduction Accumulation of β‐amyloid is a pathological hallmark of Alzheimer's disease (AD). β‐Amyloid activates the plasma contact system leading to kallikrein‐mediated cleavage of intact high‐molecular‐weight kininogen (HKi) to cleaved high‐molecular‐weight kininogen (HKc). Increased HKi cleavage is observed in plasma of AD patients and mouse models by Western blot. For potential diagnostic purposes, a more quantitative method that can measure HKc levels in plasma with high sensitivity and specificity is needed. Methods HKi/c, HKi, and HKc monoclonal antibodies were screened from hybridomas using direct ELISA with a fluorescent substrate. Results We generated monoclonal antibodies recognizing HKi or HKc specifically and developed sandwich ELISAs that can quantitatively detect HKi and HKc levels in human. These new assays show that decreased HKi and increased HKc levels in AD plasma correlate with dementia and neuritic plaque scores. Discussion High levels of plasma HKc could be used as an innovative biomarker for AD.

Keywords

• Alzheimer's disease
• Contact system activation
• HMW kininogen
• Cleaved HMW kininogen
• Plasma biomarker
• Sandwich ELISA