CAJ | 학술논문

目的：通过优化生产工艺来控制以VERO细胞为载体的狂犬疫苗生产中的残留DNA。方法采用双抗体夹心ELISA法检测评估抗原回收率，通过DNA探针杂交法检测残留DNA。采用检测抗原回收率和DNA残留率对狂犬疫苗生产中的核心环节－超滤的3个关键因素：超滤膜包的选择、超滤压力以及浓缩倍数进行评估，并评价鱼精蛋白预处理超滤液对于上述两指标的影响。结果与结论综合抗原回收率、DNA去除率以及生产层面的因素，获得了超滤工艺的最优条件，即以截留相对分子质量为7．5×105的超滤膜包、20倍浓缩和15 psi的超滤压力作为超滤工艺的核心参数。通过鱼精蛋白的预处理，初步探明超滤截留是通过分子筛的模式截留DNA，而鱼精蛋白的加入可以结合碎片化的DNA，形成较大的分子片段，从而更利于超滤截留。
목적：통과우화생산공예래공제이VERO세포위재체적광견역묘생산중적잔류DNA。방법채용쌍항체협심ELISA법검측평고항원회수솔，통과DNA탐침잡교법검측잔류DNA。채용검측항원회수솔화DNA잔류솔대광견역묘생산중적핵심배절－초려적3개관건인소：초려막포적선택、초려압력이급농축배수진행평고，병평개어정단백예처리초려액대우상술량지표적영향。결과여결론종합항원회수솔、DNA거제솔이급생산층면적인소，획득료초려공예적최우조건，즉이절류상대분자질량위7．5×105적초려막포、20배농축화15 psi적초려압력작위초려공예적핵심삼수。통과어정단백적예처리，초보탐명초려절류시통과분자사적모식절류DNA，이어정단백적가입가이결합쇄편화적DNA，형성교대적분자편단，종이경리우초려절류。
Objective To control residual DNA by optimizing methodology during the production of rabies vaccine using Vero cells as a vector .Methods The antigen recovery rate was assessed by linked immunosorbent assay-sandwich technique while the residual DNA was detected by DNA probe hybridization method .Antigen recovery and removal of DNA were the main indexes for evaluateing ultrafiltration , the vital part of rabies vaccine production .Three key factors in ultrafiltration were assessed: selection of membrane packages , ultrafiltration pressure and the concentration ratio .Then protamine was used to pretreat ultrafiltrates .Based on the two indicators mentioned above , the effect of protamine pretreat-ment on the ultrafiltrate was evaluated .Results and Conclusion The optimum condition of ultrafiltration was obtained on the basis of the general antigen recovery rate , DNA removal rate and actual production .The primary parameters of ultrafil-tration were as follows:7.5 ×105 ultrafiltration membrane packages, 20 times concentrated, 15 psi ultrafiltration pressure. After pretreatment with protamine , ultrafiltration has proved to be a molecular sieve in intercepting DNA ,while protamine can tangle the fragmented DNA and form a larger molecular segment , which is believed to be more conducive to ultrafiltra-tion interception .