Preparation Of Immunological Starch Microspheres

Liver specific lipoprotein(LSP)plays certain functions in the immunopathogenesis ofhepatitis B. It is the major antigen of hepatitis B. In this paper, the antibody against LSP waswas cross-linked with starch microspheres (SM) to gain the immunological starchmicrospheres. Immunological starch microspheres that gained the ability of targeting to liverthrough the special response between LSP and antibody against LSP can protect Liver cellsand reduce the liver damage of Hepatitis B patient.It can also carriy the medicines and makethem enriched in pathological change spot.Thus, the studies provided reference to exploringnew treatment of hepatitis B. The results were as the follows:LSP was extracted by sucrose density gradient centrifugation from fresh rat liver. Theantibody against LSP were produced by immunizing rabbits with LSP as antigen. Starchmicrospheres (SM) were prepared with inverse microemulsion. The SM activated withepichlorohydrin was cross-linked with antibody against LSP to gain the liver-targetingimmunological SM.The antibody against LSP was specific to LSP proved by direct fluorescence analysis(DFA) testing on frozen section of rat liver in which antibody against LSP was labelling withfluorescein isothiocyanate (FITC).We investigated influences of the actived process with Epoxy density(EPD) as the target.The result showed that temperature, time, concentration of NaOH and the used amount ofepichlorohydrin had great influences on EPD. The concrete results were:1. In initial periodof response, EPD increased with the elevated temperature and increased of reaction time,after the response carried on certain time, EPD started to reduce.Especially when the reactiontemperature was higher, the decreasing trendency of EDP was more obvious. In30℃responsed5h, EPD achieved maximum value.2. EPD first increased and then decreased withthe increased of reaction concentration of NaOH, and reached the maximum value when theconcentration of NaOH was2mol/L.3. In a certain range, the more epichlorohydrin in thereaction system made the higher value of EPD. EPD reached the maximum value when the amount of epichlorohydrin was0.35mL/g SM. If continuing to increase the amount ofepichlorohydrin, EPD could not increase.We studied the influences of cross-linking condition by single-factor experimemts.Optimum cross-linked pH range was9.4~9.7, optimum cross-linked time was17~20h. Theamount of antibody against LSP cross-linked to unitage SM was7.7mg/g under the optimumcross-linked condition.LSP in vitro specific binded to the unitage immunological SM was up to0.5mg/g thatshowed a high specific binding activity of antibody against LSP cross-linked on SM. Itindicated that immunological SM gained the ability of active targeting to liver, alsoilluminated that after antibody and starch microballoons cross-linking, the antibody still hadthe ability of combining antigen.