CRM197-grafted liposomes containing cardiolipin (CL) (CRM197/CL-liposomes) were used to enhance the permeability of neuron growth factor (NGF) across the blood-brain barrier (BBB) for promoting the neuroprotective effect of NGF. CRM197/CL-liposoms were incubated with a monolayer of human astrocyte (HA)-regulated human brain-microvascular endothelial cells (HBMECs) and employed to rescue SK-N-MC cells with insult of fibrillar β-amyloid peptide (1-42) (Aβ1-42). An increase in the CL mole percentage enhanced the particle size, absolute value of zeta potential, NGF entrapment efficiency, CRM197 grafting efficiency, viability of HBMECs, HAs, and SK-N-MC cells, and BBB permeability of propidium iodide (PI) and NGF, and reduced the transendothelial electrical resistance (TEER). In addition, an increase in the CRM197 weight percentage increased the particle size, absolute value of zeta potential, viability of HBMECs and HAs, and BBB permeability of PI and NGF, and decreased the CRM197 grafting efficiency and TEER. CRM197/CL-liposomes have the ability to target the BBB and to reduce neurotoxicity of Aβ142 and can be promising formulations for treating Alzheimer's disease in future medicinal application.