Study Of Split Influenza Virus Vaccine

Influenza viral inactivation vaccine is popular used in domesticat present.This vaccine contains internal and exterior antigen andlipide ,so adverse reaction comparatively large and refer to be usedcrowd above 12,immunization effect good. Split vaccine to beclearaged by split agent conserve internal and exterior antigen andadverse reatction is comparatively small and has good immunizationeffect .It is very refer to be used child and aged people.Themanufacture of spit vaccine criticality depends on the election ofsplit agent and suitable operational condition.There are threerequirements shoud be considered ①vaccine must be safe,even if itis used in immunologic injury people.②vaccine should has goodimmunization effect that it can induced completely defense effectto aim directly attack of wild strain after inoculating.③costprice of vaccine should be low .Influenza virus is RNA virus whichcomposed of nucleonic acid and cell membrane and relative molecularmass is 5900-6300.The glucoprotein ecphyma of flu virus containstwo forms that one is hemagglutinin(HA) and the other isneuramidinase (NA).All this surface glucoprotein special HAdetermines antigenic specificity of subtype and is main determinantgroup of flu virus toxicity and also is major component to induceneutralizing antibody after infection or immunization.Flu virus relative molecular mass is big ,so adverse reaction oftotivirus inactivation vaccine is big.After dealing with splitagent the relative molecular mass shrinks and adverse reaction alsodegrades large extent. In the manufacture of vaccine theimmunization effect will be affected if the split agent or splitcondition is unsuitable and cost price will be increased,so it isdisadvantage to spread use vaccine and prophylaxis flu.Ourexperimental result indicate that split effect is optimization andhemagglutinin potency has no significant difference with no splitand the yield is high.This result is important to produce largescale flu virus vaccine .The main work about our experiment are as fellows:(1) The choose of split agent: the success of multiple usageTtitonX-100 to clearage flu virus indicates that it isfeasible and craft ripe in using TritonX-100 to clearagesplit vaccine and the products is safe.(2) The research of the optimum condition in clearage flu viriusby using TritonX-100:1%TritonX-100 as final concentrationmay clearage flu virus by vibrating 1h at room temperature .(3) The scavenging of split agent: TritonX-100 and formaldehydeare scavenged by ten thousand MW ultrafiltrationmembrane.the sample obtained after column chromatography andscavenging split agent are detected HA concentration bysingle diffusion test respectively and also calculateantigen recovery rate.The recovery rate of virus exceed 96%after purifying by ten thousand MW column chromatography.(4) The establishment of detection method for remnant splitagent :Remnant Triton X-100 <500ug/ml is qualified bychromatometry.(5) The establishment of flu virus split vaccine craft flow-sheet:flu virus inoculate chick embryo → havest chick embryoallantoic fluid concentrate→purify →inactivate→purify second time→get rid of bacterium→assay monotypeviru raw liquor → prepare semi-manufacture → assaysemi-manufcture goods →subpackage → assay finalproduct.Flu virus inactivated vaccine manufactured by our group notonly fill up blank in this field in domestic but also terminatethe history that can not prduce this vaccine by our country andhave to depend on import.Further more,it will be applyextensively for its high level antibody and gently adversereaction just same as to import vaccine.