【目的】监测当前湖南省猪圆环病毒2型(Porcine circovirus type 2,PCV2)流行毒株及其衣壳蛋白(capsid protein,Cap)变异情况,并预测Cap蛋白细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)表位,为新型PCV2疫苗研制和病毒净化提供参考...【目的】监测当前湖南省猪圆环病毒2型(Porcine circovirus type 2,PCV2)流行毒株及其衣壳蛋白(capsid protein,Cap)变异情况,并预测Cap蛋白细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)表位,为新型PCV2疫苗研制和病毒净化提供参考依据。【方法】本研究对2019-2021年于湖南省6个地区收集的17份PCV2阳性组织样品进行PCV2全基因组序列扩增及测序分析,绘制系统进化树,利用生物信息学方法分析Cap蛋白氨基酸变异情况,并预测CTL表位。【结果】系统进化树显示,获得的17株PCV2全基因组序列中,1株PCV2a、7株PCV2b和9株PCV2d。Cap蛋白氨基酸序列比对分析发现,共有16个氨基酸残基突变位点位于病毒Cap蛋白表面,且有11个突变位点参与构象型表位的形成。此外,共预测出9个PCV2 Cap蛋白潜在的CTL表位,其中4个表位(16-24、28-36、136-144和179-187位氨基酸)在GenBank的1610株PCV2不同基因型毒株中高度保守。通过TCR-pMHC复合物3D结构对4个保守性表位进一步分析,结果显示,这4个肽段均能与MHCⅠ分子和TCR形成稳定的TCR-pMHC复合物。【结论】本研究结果表明,PCV2b和PCV2d为当前湖南省主要流行的基因型,且表现出高度变异;预测的CTL表位可作为候选抗原表位。展开更多
Objective This study aimed to design and optimize the water extraction process for Chuantieling(喘贴灵,CTL)based on the concept of quality by design(QbD).Methods The single-factor experiments were used to select the b...Objective This study aimed to design and optimize the water extraction process for Chuantieling(喘贴灵,CTL)based on the concept of quality by design(QbD).Methods The single-factor experiments were used to select the best experimental points of CTL water extraction.On this basis,the transfer rate of ephedrine hydrochloride and sin-apine thiocyanate,and the yield of the extract were evaluated as the evaluation indicators.The liquid-solid ratio,extraction time,and pH value were selected as the main factors to carry out the Box-Behnken design(BBD).A mathematical model of the critical process parameters(CPPs)and critical quality attributes(CQAs)was established,the interaction between CQAs and CPPs was examined,and the CTL extraction process design space was established and optimized,which guaranteed the stability of the process.The high performance liquid chro-matography(HPLC)was used to analyze the main active compounds in the CTL extract.Results Through single-factor experiments,the best experimental parameters were found to be a liquid-solid ratio of 8∶1,extraction time of 90 min,pH value of 4,and extraction times of three.The experimental variance analysis results of the BBD showed that the P value of the re-gression model was less than 0.05,and the lack-of-fit value was greater than 0.01,indicating that the constructed model had good predictive ability.The operating space of the CPPs of the CTL water extraction process was combined with the actual production situation.In the pro-duction situation,the best extraction process was nine times of water addition,72 min of ex-traction time,and 4.5 of pH value.The HPLC results showed that the peak areas of ephedrine hydrochloride,sinapine thiocyanate,tetrahydropalmatine,methyl eugenol,cinnamaldehyde,and 6-gingerol in the CTL extract accounted for 0.94%,14.32%,0.78%,31.23%,0.34%,and 0.44%of the total peak area,respectively.Conclusion The water extraction process design space of CTL based on QbD was conducive to actual production operations,ensuring the stability of the process.展开更多
为简化细胞病毒T细胞(cytotoxicity T lymphocytek,CTL)表位鉴定方法,应用改进的人工神经网络方法定量研究了短肽与MHC(major histocompatibility complex)分子结合亲合力的关系,并建立了CTL表位的预测模型,得到了预测模型最优性能参数...为简化细胞病毒T细胞(cytotoxicity T lymphocytek,CTL)表位鉴定方法,应用改进的人工神经网络方法定量研究了短肽与MHC(major histocompatibility complex)分子结合亲合力的关系,并建立了CTL表位的预测模型,得到了预测模型最优性能参数.用此模型对短肽与HLA-A*0201分子结合的805个预测样本进行了预测,预测准确度达到73.8%.对来自黑色素MAGE-2的短肽与MHC分子的结合亲合力也进行了预测,结果较好.展开更多
文摘【目的】监测当前湖南省猪圆环病毒2型(Porcine circovirus type 2,PCV2)流行毒株及其衣壳蛋白(capsid protein,Cap)变异情况,并预测Cap蛋白细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)表位,为新型PCV2疫苗研制和病毒净化提供参考依据。【方法】本研究对2019-2021年于湖南省6个地区收集的17份PCV2阳性组织样品进行PCV2全基因组序列扩增及测序分析,绘制系统进化树,利用生物信息学方法分析Cap蛋白氨基酸变异情况,并预测CTL表位。【结果】系统进化树显示,获得的17株PCV2全基因组序列中,1株PCV2a、7株PCV2b和9株PCV2d。Cap蛋白氨基酸序列比对分析发现,共有16个氨基酸残基突变位点位于病毒Cap蛋白表面,且有11个突变位点参与构象型表位的形成。此外,共预测出9个PCV2 Cap蛋白潜在的CTL表位,其中4个表位(16-24、28-36、136-144和179-187位氨基酸)在GenBank的1610株PCV2不同基因型毒株中高度保守。通过TCR-pMHC复合物3D结构对4个保守性表位进一步分析,结果显示,这4个肽段均能与MHCⅠ分子和TCR形成稳定的TCR-pMHC复合物。【结论】本研究结果表明,PCV2b和PCV2d为当前湖南省主要流行的基因型,且表现出高度变异;预测的CTL表位可作为候选抗原表位。
基金Hunan University of Traditional Chinese Medicine Graduate Training Quality Engineering Project(2019CX57)the First-class Discipline Project on Chinese Pharmacology of Hunan University of Chinese Medicine (201803)。
文摘Objective This study aimed to design and optimize the water extraction process for Chuantieling(喘贴灵,CTL)based on the concept of quality by design(QbD).Methods The single-factor experiments were used to select the best experimental points of CTL water extraction.On this basis,the transfer rate of ephedrine hydrochloride and sin-apine thiocyanate,and the yield of the extract were evaluated as the evaluation indicators.The liquid-solid ratio,extraction time,and pH value were selected as the main factors to carry out the Box-Behnken design(BBD).A mathematical model of the critical process parameters(CPPs)and critical quality attributes(CQAs)was established,the interaction between CQAs and CPPs was examined,and the CTL extraction process design space was established and optimized,which guaranteed the stability of the process.The high performance liquid chro-matography(HPLC)was used to analyze the main active compounds in the CTL extract.Results Through single-factor experiments,the best experimental parameters were found to be a liquid-solid ratio of 8∶1,extraction time of 90 min,pH value of 4,and extraction times of three.The experimental variance analysis results of the BBD showed that the P value of the re-gression model was less than 0.05,and the lack-of-fit value was greater than 0.01,indicating that the constructed model had good predictive ability.The operating space of the CPPs of the CTL water extraction process was combined with the actual production situation.In the pro-duction situation,the best extraction process was nine times of water addition,72 min of ex-traction time,and 4.5 of pH value.The HPLC results showed that the peak areas of ephedrine hydrochloride,sinapine thiocyanate,tetrahydropalmatine,methyl eugenol,cinnamaldehyde,and 6-gingerol in the CTL extract accounted for 0.94%,14.32%,0.78%,31.23%,0.34%,and 0.44%of the total peak area,respectively.Conclusion The water extraction process design space of CTL based on QbD was conducive to actual production operations,ensuring the stability of the process.
文摘为简化细胞病毒T细胞(cytotoxicity T lymphocytek,CTL)表位鉴定方法,应用改进的人工神经网络方法定量研究了短肽与MHC(major histocompatibility complex)分子结合亲合力的关系,并建立了CTL表位的预测模型,得到了预测模型最优性能参数.用此模型对短肽与HLA-A*0201分子结合的805个预测样本进行了预测,预测准确度达到73.8%.对来自黑色素MAGE-2的短肽与MHC分子的结合亲合力也进行了预测,结果较好.
