[Preparation and preliminary evaluation of KGDS-targeted ultrasound contrast agent].
OBJECTIVE
To prepare a thrombus-targeted ultrasonic contrast agent and to investigate its targeted ability to fresh blood clots.


METHODS
We first synthesized FITC-KGDS-Palm compound, and then prepared thrombus-targeted microbubbles using "ultrasound & high speed shearing method". Fluorescence labeling thrombus-specific peptides and KGDS, directed at the activated glycoprotein(GP)IIb/IIIa receptor of platelets were attached to the surface of lipid microbubbles. The concentration and size of TUCA were measured by Malvern Zeta Sizer Nano-ZS590 and Coulter counter. Immunofluorescence was applied to confirm the conjugation. The conjunct ratio was assessed by flow cytometer (FCM).


RESULTS
The KGDS-TUCA was straw yellow turbid liquor, and the concentration was 1.5 x 10(9)/mL, and the average size was 1.5 microm. The targeted microbubbles conjugated with the thrombus-specific peptides showed bright green rings by fluorescence microscope. FCM demonstrated that the wavelength of shell of KGDS-TUCA changed greatly, and the conjunct ratio was 90.04%. In vitro study showed KGDS-TUCA remained stable for 48 h at 4 degree C and target-attached to blood clots and showed good stability.


CONCLUSIONS
The ultrasound & high speed shearing method to prepare TUCA is easy and in favor of purification. KGDS-TUCA has high specific biological activity. The conjunct ratio and stability of KGDS-TUCA are excellent.
Gao
Feng
F
Institute of Cell Transplantation & Gene Therapy, Third Xiangya Hospital, Central South University, Changsha 410013, China.
Ding
Yanfei
Y
Sheng
Xiaoxi
X
Wang
Wei
W
Liang
Qi
Q
Luo
Zhuoqiong
Z
Zhou
Ping
P
Li
Hui
H
D000328:Adult; D000596:Amino Acids / Q000737:chemistry; D003287:Contrast Media / Q000138:chemical synthesis*; D006801:Humans; D045423:Microbubbles; D057054:Molecular Imaging / Q000379:methods*; D015335:Molecular Probes; D010455:Peptides / Q000737:chemistry; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism; D012680:Sensitivity and Specificity; D013927:Thrombosis / Q000000981:diagnostic imaging*; D014463:Ultrasonography
D000596:Amino Acids; D003287:Contrast Media; D015335:Molecular Probes; D010455:Peptides; D019039:Platelet Glycoprotein GPIIb-IIIa Complex