The Fabrication Of Anti-PSA-SPIO And Anti-HER-2/neu-SPIO With Which Molecular Imaging In LNCaP And PC-3 Were Invested

Objective:1. To fabrication the anti-PSA-SPIO and anti-HER-2/neu-SPIO.To test their diameter and shape. To evaluate the binding rate of antibody and SPIO.2. To cultivate LNCaP and PC-3.To examine the expression of PSA and HER-2/neu in the two cells with immunocytochemistry.3. To analysis the target effects of antibody contrast agents to LNCaP and PC-3 cells with electron microscope ,flow cytometry and MR.Methods:1. First, The anti-PSA-SPIO and anti-HER-2/neu-SPIO was fabricated with chemical method. second, we tested their diameter and shape.third, To evaluate the binding rate of antibody and SPIO with Folin-phenol method.2. To cultivate LNCaP and PC-3.To examine the expression of PSA and HER-2/neu in the two cells with immunocytochemistry method.3. Five samples (anti-PSA,anti-PSA-SPIO,anti-HER-2/neu,anti-HER-2/neu– SPIO,SPIO) were assayed with LNCaP and PC-3 by flow cytometry(FCM) in order to evaluate the immunoactivity respectively.4. We tested the activity and the circumstance in cells of McAb-SPIO and analysised the target effects of antibody contrast agents.Results:1. The results of assaying protein level and electron microscope proved that the two McAb was conjugated with SPIO effectively.The conjuction rate of anti-PSA and SPIO was 96.5%. The conjuction rate of anti-HER-2/neu and SPIO was 96.5%.The diameters of particles were between 50and 80nm.2. Concluded from immunocytochemistry test , LNCaP expressed PSA but HER-2/neu,PC-3 expressed HER-2/neu but PSA.3. Concluded from flow cytometry results, LNCaP combined with anti-PSA and anti-PSA-SPIO in a high rate , PC-3 combined with anti-HER-2/neu and anti-HER-2/neu-SPIO in a high rate too.4. Investigated from electron microscope ,anti-PSA-SPIO get into LNCaP cells more easily than other, anti-HER-2/neu-SPIO get into PC-3 cells more easily too.5. Tested by MR,the signal of the two samples (LNCap and anti-PSA-SPIO, PC-3 and anti-HER-2/neu-SPIO) turned down sharply inT2WI.Conclusion:The anti-PSA-SPIO and anti-HER-2/neu-SPIO was fabricated successfully. The anti-PSA-SPIO targeted to LNCaP. The anti-HER-2/neu-SPIO targeted to PC-3. The two probes all had novel physiochemical property and high level of immunoactivity.They all had targeted enhanced effect to cells in magnetic resonance scanning. Both of them will improve the diagnosis of either hormonal dependent PCa or hormonal independent PCa.Its successfully preparation will provide us a new credible technical method in molecular imaging of tumor .