CAJ | 학술논문

目的：对重组表达HPV68 b L1蛋白的汉逊酵母工程菌进行高密度发酵，并对L1蛋白形成的类病毒颗粒（virus-like particles，VLPs）进行纯化、鉴定及免疫学效果评价。方法 HPV68b L1的酵母工程菌经过发酵，系列纯化和颗粒重构后获得VLPs抗原，对该抗原进行蛋白纯度、颗粒形态及免疫原性等检定，与铝佐剂吸附后免疫动物，利用假病毒体外中和试验评价其对HPV68 b型及HPV68a型的保护效果。结果发酵后的工程菌经过纯化和颗粒重构后，获得纯度大于95％的VLPs，电镜结果显示，其颗粒形态均一，大小与天然病毒颗粒接近。假病毒体外中和试验结果显示， HPV68b型VLPs抗原可诱导小鼠体内产生较高滴度的中和抗体，同时对HPV68a型也具有一定的交叉保护作用。结论验证了重组HPV68 b型VLPs蛋白用于疫苗生产的可行性，可作为多价HPV疫苗的组分。
목적：대중조표체HPV68 b L1단백적한손효모공정균진행고밀도발효，병대L1단백형성적류병독과립（virus-like particles，VLPs）진행순화、감정급면역학효과평개。방법 HPV68b L1적효모공정균경과발효，계렬순화화과립중구후획득VLPs항원，대해항원진행단백순도、과립형태급면역원성등검정，여려좌제흡부후면역동물，이용가병독체외중화시험평개기대HPV68 b형급HPV68a형적보호효과。결과발효후적공정균경과순화화과립중구후，획득순도대우95％적VLPs，전경결과현시，기과립형태균일，대소여천연병독과립접근。가병독체외중화시험결과현시， HPV68b형VLPs항원가유도소서체내산생교고적도적중화항체，동시대HPV68a형야구유일정적교차보호작용。결론험증료중조HPV68 b형VLPs단백용우역묘생산적가행성，가작위다개HPV역묘적조분。
Objective To evaluate the immunogenicity of the recombinant human papillomavirus type 68b (HPV68b) virus-like particles(VLPs)in a mouse model.Methods The L1 protein of HPV type 68b was successful expressed in the Hansenula polymorpha strain (NVSI-68b-1).Processes including purifi-cation and reconstitution were performed to achieve pure HPV 68b VLPs.The purity, morphology and immu-nogenicity of the purified HPV 68 b VLPs were further analyzed .The BALB/c mice were immunized with HPV68b VLPs formulated on aluminum adjuvant .Pseudovirus-neutralizing antibody ( PsV NAb) assay was performed to detect the neutralizing antibodies in serum samples .Results The HPV 68 b L1 VLPs were ob-tained as indicated by the results of SDS-PAGE, Western blot assay , HPLC, electron microscopy and dy-namic light scattering with a high purity of 95%.Transmission electron microscopy and dynamic light scat-tering analysis revealed that the HPV68b L1 VLPs resembled the native virus with an average particle diame-ter of 50 nm.High levels of HPV68b-neutralizing antibodies were detected in serum samples from the mice immunized with HPV68b L1 VLPs.Moreover, a cross-protective efficacy of HPV68b L1 VLPs for HPV68a was observed .Conclusion This study suggested that the recombinant HPV 68 b VLPs expressed in a Han-senula polymorpha strain might be used as a potential candidate for the development of HPV vaccine .