Preparation and targeting study in vitro of P-selectin antibody loaded targeted ultrasound contrast agent
10.3760/cma.j.issn.1004-4477.2013.04.021
- VernacularTitle:携抗P-选择素单抗靶向超声造影剂的制备及体外寻靶实验研究
- Author:
Xinzhao LIU
;
Jianli REN
;
Zhigang WANG
- Publication Type:Journal Article
- Keywords:
Microbubbles;
P-selectin
- From:
Chinese Journal of Ultrasonography
2013;(4):349-354
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare a kind of P-selectin antibody loaded-targeted ultrasound contrast agent,and to investigate its ability of targeting in vitro.Methods P-selectin antibody loaded-targeted ultrasound contrast agent was prepared via an avidin-biotin bridge.Its basic property was determined.Human umbilical vein endothelial cells (HUVECs) were stimulated to express P-selectin by applying different doses of recombinant human interleukin-4 (rhIL-4) and histamine.The expression level of P-selectin was detected by immunofluorescence technique,and explored rhIL-4 optimal stimulation dose.The experiment group was divided into 3 groups,including targeted ultrasound contrast agent,isotype control ultrasound contrast agent and blank ultrasound contrast agent.Targeting abilities of 3 groups were observed by adhering to treated HUVECs and untreated HUVECs,respectively.Results P-selectin antibody loaded targeted ultrasound contrast agent was prepared successfully.The average diameter was (2.24 ± 0.71)μm.The average Zeta potential was (-2.75 ± 0.84)mV.The concentration was (3.0 ± 0.3) × 109/ml.The rate of antibody binding was as high as 99.80％.RhIL-4 and histamine could stimulate HUVECs to express P-selectin,and optimal stimulation dose was obtained.Targeting experiment showed that targeted ultrasound contrast agent could preferably adhere to HUVECs stimulated by optimal dose of rhIL-4 and histamine.Compared with other groups,there were significant differences.Conclusions P-selectin antibody loaded targeted ultrasound contrast agent was prepared successfully via an avidin-biotin bridge.The targeted ultrasound contrast agent could effectively adhere to HUVECs stimulated by rhIL-4 and histamine.
