Preparation Of Long Circulating Immunoliposomes With The Specific Single Chain Antibody Against Schistosoma Japonicum SIEA26-28 Ku And Praziquantel

ObjectiveTo prepare the long circulating immunoliposomes wrapping Praziquantel against Schistosoma japonicum as the preliminary work of the targeted therapy.Methods1,Expression and purification of SIEA26-28 ku-scFvE.coli BL21 (DE3) containing plasmid pET32a/SIEA26-28 ku-scFv was induced by IPTG. The expressed proteins were purified by Ni2+ column affinity chromatography. The concentration of purified proteins was determined. The purified proteins were identified by SDS-PAGE and Western blot.2,Preparation of the long circulating liposomes wrapping praziquantelThe long circulating liposomes wrapping praziquantel were prepared by the method of thin film evaporation and Ultrasonic dispersion.And its envelopment efficiency was determined.3,Preparation of the long circulating immunoliposomes with SIEA26-28KU-scFvBy the new shaped bifunctional crosslinker SPDP, SIEA26-28 ku-scFv was connected to liposomes in order to construct the third generation of long circulating immunoliposomes wrapping praziquantel. And its coupling rate was calculated.Results1,SIEA26-28 ku-scFv was highly expressed.The proteins were purified by Ni2+column affinity chromatography. Western blot showed that SIEA26-28 scFv can specifically recognize SIEA.2,The Long circulating liposomes wrapping praziquantel were prepared successfully.And its envelopment efficiency was 56.1%.3,The third generation of long circulating immunoliposomes wrapping praziquantel that coupled with SIEA26-28 ku-scFv were constructed successfully. And its coupling rate was 21.3%.conclusionSIEA26-28 ku-scFv was extracted and purified successfully. The third generation of long circulating immunoliposomes with SIEA26-28 ku-scFv and Praziquantel were prepared,which laid the basic work of the targeted therapy against Schistosoma japonicum for the next step.