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Antigen epitopes of animal coronaviruses:a mini-review
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作者 Mingjun Su Guanghui Zheng +1 位作者 Xiangwen Xu Houhui Song 《Animal Diseases》 CAS 2024年第1期19-26,共8页
Coronaviruses are widespread in nature and can infect mammals and poultry,making them a public health concern.Globally,prevention and control of emerging and re-emerging animal coronaviruses is a great challenge.The m... Coronaviruses are widespread in nature and can infect mammals and poultry,making them a public health concern.Globally,prevention and control of emerging and re-emerging animal coronaviruses is a great challenge.The mecha-nisms of virus-mediated immune responses have important implications for research on virus prevention and control.The antigenic epitope is a chemical group capable of stimulating the production of antibodies or sensitized lympho-cytes,playing an important role in antiviral immune responses.Thus,it can shed light on the development of diagnos-tic methods and novel vaccines.Here,we have reviewed advances in animal coronavirus antigenic epitope research,aiming to provide a reference for the prevention and control of animal and human coronaviruses. 展开更多
关键词 Animal coronavirus Antigen epitope b-cell epitope T-cell epitope Immune responses Vaccines
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Identification of an HLA-A~* 0201-restricted CD8^+ T-cell epitope SSp-1 of SARS-CoV spike protein 被引量:21
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作者 WangB ChenH JiangX ZhangM WanT LiN ZhouX WuY YangF YuY WangX YangR CaoX 《第二军医大学学报》 CAS CSCD 北大核心 2004年第9期969-969,共1页
A novel coronavirus, severe acute respiratory syndrome (SA RS)-associated coronavirus (SARS-CoV), has been identified as the causal agent of SARS. Spike (S) protein is a major structural glycoprotein of the SARS virus... A novel coronavirus, severe acute respiratory syndrome (SA RS)-associated coronavirus (SARS-CoV), has been identified as the causal agent of SARS. Spike (S) protein is a major structural glycoprotein of the SARS virus and a potential target for SARS-specific cell-mediated immune responses. A pa nel of S protein-derived peptides was tested for their binding affinity to HLA -A *0201 molecules. Peptides with high affinity for HLA-A *0201 were then as se ssed for their capacity to elicit specific immune responses mediated by cytotoxi c T lymphocytes (CTLs) both in vivo, in HLA-A2.1/K b transgenic mice, a nd in vitro, from peripheral blood lymphocytes (PBLs) harvested from healthy HLA-A 2.1 + donors. SARS-CoV protein-derived peptide-1 (SSp-1 RLNEVAKNL), induced pepti de-specific CTLs both in vivo (transgenic mice) and in vitro (human PBL s), which specifically released interferon-gamma (IFN-gamma) upon stimulation with SSp-1-pulsed autologous dendritic cells (DCs) or T2 cells. SSp-1-specif ic CTLs also lysed major histocompatibility complex (MHC)-matched tumor cell lines engineered to express S proteins. HLA-A *0201-SSp-1 tetramer staining re vealed the presence of significant populations of SSp-1-specific CTLs in SSp- 1-induced CD8 + T cells. We propose that the newly identified epitope SSp-1 w ill help in the characterization of virus control mechanisms and immunopathology in SARS-CoV infection, and may be relevant to the development of immunotherape utic approaches for SARS. 展开更多
关键词 SARS-CoV spike protein T-cell epitope SSp-1 of identification of an HLA-A restricted CD8 HLA cell CD
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Identification of an HLA-A*0201-restricted CD8^+ T-cell epitope SSp-1 of SARS-CoV spike protein
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作者 WangB ChenH JiangX ZhangM WanT LiN ZhouX WuY YangF YuY WangX YangR CaoX 《第二军医大学学报》 CAS CSCD 北大核心 2005年第7期794-794,共1页
A novel coronavirus, severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV), has been identified as the causal agent of SARS. Spike (S) protein is a major structural glycoprotein of the SARS virus ... A novel coronavirus, severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV), has been identified as the causal agent of SARS. Spike (S) protein is a major structural glycoprotein of the SARS virus and a potential target for SARS-specific cell-mediated immune responses. A panel of S protein-derived peptides was tested for their binding affinity to HLA-A*0201 molecules. Peptides with high affinity for HLA-A*0201 were then assessed for their capacity to elicit specific immune responses mediated by cytotoxic T lymphocytes (CTLs) both in vivo, in HLA-A2.1/K b transgenic mice, and in vitro, from peripheral blood lymphocytes (PBLs) harvested from healthy HLA-A2.1(+) donors. SARS-CoV protein-derived peptide-1 (SSp-1 RLNEVAKNL), induced peptide-specific CTLs both in vivo (transgenic mice) and in vitro (human PBLs), which specifically released interferon-gamma (IFN-γ) upon stimulation with SSp-1-pulsed autologous dendritic cells (DCs) or T2 cells. SSp-1-specific CTLs also lysed major histocompatibility complex (MHC)-matched tumor cell lines engineered to express S proteins. HLA-A*0201-SSp-1 tetramer staining revealed the presence of significant populations of SSp-1-specific CTLs in SSp-1-induced CD8+ T cells. We propose that the newly identified epitope SSp-1 will help in the characterization of virus control mechanisms and immunopathology in SARS-CoV infection, and may be relevant to the development of immunotherapeutic approaches for SARS. 展开更多
关键词 SARS identification of an HLA-A*0201-restricted CD8 T-cell epitope SSp-1 of SARS-CoV spike protein HLA cell CD
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Bioinformatics analysis of the structure and linear B-cell epitopes of aquaporin-3 from Schistosoma japonicum 被引量:11
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作者 Jie Song Qing-Feng He 《Asian Pacific Journal of Tropical Medicine》 SCIE CAS 2012年第2期107-109,共3页
Objective:To analyze the structure of aquaporins-3(AQP-3) from Schistosoma japonicum(SJAQP-3) using bioinformalical methods,and to provid of references for vaccine targets research.Methods:Protparam,BepiPred,TMHMM Ser... Objective:To analyze the structure of aquaporins-3(AQP-3) from Schistosoma japonicum(SJAQP-3) using bioinformalical methods,and to provid of references for vaccine targets research.Methods:Protparam,BepiPred,TMHMM Server,MLRC,Geno3d,DNA star software packages were used to predict the physical and chemical properties,hydrophilicity plot, flexibility regions,antigenic index,surface probability plot,secondary structure,and tertiary structure of amino acid sequence of SJAQP-3.Results:SJAQP-3 had six transmembrane regions and two half-spanning helices that form a central channel.The half-spanning helices fold into the centre of the channel.Either of the half-spanning helix had a conserved motif of NPA common to all aquaporins.Predicted linear B-Cell epitopes were most likely at the N-terminal amino acid residues of Saa-7aa,59aa- 62aa,225aa-230aa,282aa -288aa,294aa -29Saa and 305aa -307aa area.59aa- 62aa,22Saa-230aa located outside the membrane,the others located inside the cell.Conclusions:SJAQP-3 is a integral membrane protein in Schistosoma japonicum tegument.There are six potential epitopes in SJ AQP-3.It might be a potential molecular target for the development of vaccines. 展开更多
关键词 SCHISTOSOMA JAPONICUM Aquaporins-3 Bioinformatics LINEAR b-cell epitopes Vaccine target
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Helicobacter pylori specific immune response induced by conservative flagellin linear B-cell epitope 被引量:1
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作者 Wan-ShengJi Jia-LuHu +4 位作者 Kai-ChunWu Jun-WenQiu Zhe-YiHan JieDing Dai-MingFan 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第23期3528-3532,共5页
AIM:To testify the immunogenicity of a conservative B-cell linear epitope of Helicobacter pylori ( H pylori) flagellin A. METHODS: Different programs were used to analyze the secondary structure, molecular hydropathy,... AIM:To testify the immunogenicity of a conservative B-cell linear epitope of Helicobacter pylori ( H pylori) flagellin A. METHODS: Different programs were used to analyze the secondary structure, molecular hydropathy, and surface accessibility of Hpyloriflagellin A. Linear B-cell epitopes were estimated based on the structural and physiochemical information. Analysis of residue divergence was proposed to screen a conservative linear epitope. The 29-peptide (Pep29mer) synthesized by chemical method, including the predicted conservative B-cell epitope and a known K^2d compatible T-cell epitope, was used to immunize mice, and then H pylori-specific antibodies were detected by ELISA. RESULTS: Based on the analyses of divergent amino acid residues, structural and physiochemical characteristics, it was strongly suggested that the short fragment NDSDGR was the core of a conservative linear epitope in flagellin A. Animals immunized by Pep29mer acquired efficient immune response. In detail, serum Hpylori-specific IgA and IgGl increased significantly in immunized group, while IgG2a only had an insignificant change. Hpylori-specific IgA in gastrointestinal flushing fluid also increased significantly. CONCLUSION: The conservative short fragment NDSDGR is the core of a linear B-cell epitope of flagellin A. 展开更多
关键词 Helicobacter pylori Flagellin A b-cell epitope
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Theoretical Study of Continuous B-Cell Epitopes with Developed BP Neural Network
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作者 Yajie Cao Jinglin Liu +2 位作者 Tao Liu Dejiang Liu Yunfei Wu 《Computational Chemistry》 2016年第3期83-90,共8页
In order to identify continuous B-cell epitopes effectively and to increase the success rate of experimental identification, the modified Back Propagation artificial neural network (BP neural network) was used to pred... In order to identify continuous B-cell epitopes effectively and to increase the success rate of experimental identification, the modified Back Propagation artificial neural network (BP neural network) was used to predict the continuous B-cell epitopes, and finally the predictive model for the B-cells epitopes was established. Comparing with the other predictive models, the prediction performance of this model is more excellent (AUC = 0.723). For the purpose of verifying the performance of the model, the prediction to the SWISS PROT NUMBER: P08677 was carried on, and the satisfying results were obtained. 展开更多
关键词 Continuous b-cell epitopes BP Neural Network Theory Method Predictive Model
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An immunoglobulin Y that specifically binds to an in silico-predicted unique epitope of Zika virus non-structural 1 antigen 被引量:2
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作者 Leonardo A.Guevarra Jr Scott Dean P.De Sagon +5 位作者 Treena Rica D.Teh Maria Katrina Diana MCruz Nikki Cyrill C.Capistrano Austine James Z.StaMaria Laarni Grace M.Corales Leslie Michelle M.Dalmacio 《Asian Pacific Journal of Tropical Medicine》 SCIE CAS 2022年第1期35-43,共9页
Objective:To identify unique immunogenic epitopes of Zika virus non-structural 1(NS1)antigen and produce immunoglobulin Y(IgY)for potential use in he diagnosis of of Zika virus infection.Methods:Immunogenic epitopes w... Objective:To identify unique immunogenic epitopes of Zika virus non-structural 1(NS1)antigen and produce immunoglobulin Y(IgY)for potential use in he diagnosis of of Zika virus infection.Methods:Immunogenic epitopes were identified using in silico B-cell epitope prediction.A synthetic peptide analog of the predicted epitope was used to induce antipeptide IgY production in hens which was purified using affinity chromatography.Presence of purified IgY and its binding specificity were performed by gel electrophoresis and ELISA,respectively.Results:Out of the nine continuous epitopes identified,the sequence at position 193-208(LKVREDYSLECDPAVI)was selected and used to produce anti-peptide IgY.The produced IgY was found to bind to the synthetic analog of the Zika virus NS1 immunogenic epitope but not to other flaviviruses and random peptides from other pathogens.Conclusions:In this study,we identified an immunogenic epitope unique to Zika virus that can be used to develop a serodiagnostic tool that specifically detect Zika virus infection. 展开更多
关键词 Immunoglobulin Y IgY b-cell epitope prediction FLAVIVIRUS Non-structural 1 antigen Zika virus
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Prediction of promiscuous T-cell epitopes in the Zika virus polyprotein:An in silico approach
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作者 Hamza Dar Tahreem Zaheer +5 位作者 Muhammad Talha Rehman Amjad Ali Aneela Javed Gohar Ayub Khan Mustafeez Mujtaba Babar Yasir Waheed 《Asian Pacific Journal of Tropical Medicine》 SCIE CAS 2016年第9期822-828,共7页
Objective:To predict immunogenic promiscuous T-cell epitopes from the polyprotein of the Zika virus using a range of bioinformatics tools.To date,no epitope data are available for the Zika virus in the IEDB database.M... Objective:To predict immunogenic promiscuous T-cell epitopes from the polyprotein of the Zika virus using a range of bioinformatics tools.To date,no epitope data are available for the Zika virus in the IEDB database.Methods:We retrieved nearly 54 full length polyprotein sequences of the Zika virus from the NCBI database belonging to different outbreaks.A consensus sequence was then used to predict the promiscuous T cell epitopes that bind MHC 1 and MHC II alleles using Propred1 and Propred immunoinformatic algorithms respectively.The antigencity predicted score was also calculated for each predicted epitope using the Vaxi Jen 2.0 tool.Results:By using Pro Pred1,23 antigenic epitopes for HLA class I and 48 antigenic epitopes for HLA class II were predicted from the consensus polyprotein sequence of Zika virus.The greatest number of MHC class I binding epitopes were projected within the NS5(21%),followed by Envelope(17%).For MHC class II,greatest number of predicted epitopes were in NS5(19%) followed by the Envelope,NS1 and NS2(17% each).A variety of epitopes with good binding affinity,promiscuity and antigenicity were predicted for both the HLA classes.Conclusion:The predicted conserved promiscuous T-cell epitopes examined in this study were reported for the first time and will contribute to the imminent design of Zika virus vaccine candidates,which will be able to induce a broad range of immune responses in a heterogeneous HLA population.However,our results can be verified and employed in future efficacious vaccine formulations only after successful experimental studies. 展开更多
关键词 Zika VIRUS b-cell epitopeS T-CELL epitopeS Vaccine ANTIGENICITY
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新型冠状病毒E蛋白B细胞表位的预测及鉴定
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作者 张鹏飞 刘钧 +5 位作者 邹紫阳 康喜龙 宋丽 焦新安 孟闯 潘志明 《中国人兽共患病学报》 CAS CSCD 北大核心 2024年第9期807-813,共7页
目的利用生物信息学方法预测SARS-CoV-2 E蛋白B细胞表位,并利用小鼠多抗血清、人新型冠状病毒阳性血清等进行验证,以明确SARS-CoV-2 E蛋白的优势B细胞表位。方法使用SOPMA、Expasy、SWISS-MODEL及IEDB数据库和BepiPred-2.0等软件预测SAR... 目的利用生物信息学方法预测SARS-CoV-2 E蛋白B细胞表位,并利用小鼠多抗血清、人新型冠状病毒阳性血清等进行验证,以明确SARS-CoV-2 E蛋白的优势B细胞表位。方法使用SOPMA、Expasy、SWISS-MODEL及IEDB数据库和BepiPred-2.0等软件预测SARS-CoV-2 E蛋白的结构及B细胞表位;通过大肠杆菌系统表达并纯化GST标签重组表位蛋白片段,以Western blotting和间接ELISA方法检测表位蛋白与小鼠和人SARS-CoV-2 E蛋白阳性多抗血清的反应性,以初步鉴定SARS-CoV-2 E蛋白的B细胞表位。结果表位预测结果显示,E蛋白含有线性B细胞表位Ser6-Val14和Tyr57-Pro71,构象表位涉及的氨基酸序列为Glu8-Val14、Leu39-Tyr59、Ser60-Leu65;表达并纯化含有预测表位的E蛋白片段E1(Ser6-Val14表位)、E3(Tyr57-Pro71)以及阴性对照片段E2(不含表位序列),Western blotting和间接ELISA结果均显示抗E蛋白小鼠多抗和人新型冠状病毒阳性血清只与E1、E3蛋白片段呈阳性反应而与E2蛋白片段均为阴性反应,显示E蛋白线性B细胞表位预测正确。结论本研究成功预测并初步鉴定出SARS-CoV-2 E蛋白2个线性B细胞表位,为新型冠状病毒疫苗制备和免疫应答特性分析等提供参考。 展开更多
关键词 新型冠状病毒 B细胞表位 线性表位 预测 鉴定
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iLBE for Computational Identification of Linear B-cell Epitopes by Integrating Sequence and Evolutionary Features
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作者 Md.Mehedi Hasan Mst.Shamima Khatun Hiroyuki Kurata 《Genomics, Proteomics & Bioinformatics》 SCIE CAS CSCD 2020年第5期593-600,共8页
Linear B-cell epitopes are critically important for immunological applications,such as vaccine design,immunodiagnostic test,and antibody production,as well as disease diagnosis and therapy.The accurate identification ... Linear B-cell epitopes are critically important for immunological applications,such as vaccine design,immunodiagnostic test,and antibody production,as well as disease diagnosis and therapy.The accurate identification of linear B-cell epitopes remains challenging despite several decades of research.In this work,we have developed a novel predictor,Identification of Linear B-cell Epitope(i LBE),by integrating evolutionary and sequence-based features.The successive feature vectors were optimized by a Wilcoxon-rank sum test.Then the random forest(RF)algorithm using the optimal consecutive feature vectors was applied to predict linear B-cell epitopes.We combined the RF scores by the logistic regression to enhance the prediction accuracy.iLBE yielded an area under curve score of 0.809 on the training dataset and outperformed other prediction models on a comprehensive independent dataset.iLBE is a powerful computational tool to identify the linear B-cell epitopes and would help to develop penetrating diagnostic tests.A web application with curated datasets for iLBE is freely accessible at http://kurata14.bio.kyutech.ac.jp/iLBE/. 展开更多
关键词 Linear b-cell epitope BLAST Feature encoding Feature selection Random forest
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Identification of a Novel Universal Potential Epitope on the Cytoplasmic Tail of H7N9 Virus Hemagglutinin
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作者 Xi Liu Li Ding +8 位作者 Jing Yuan Jian Liao Lian Duan Wenfei Wang Weiguo Tan Weiye Yu Boping Zhou Xinchun Chen Zheng Yang 《Virologica Sinica》 SCIE CAS CSCD 2019年第3期334-337,共4页
Dear Editor,H7N9 is a recently identified subtype of influenza A virus that caused a major outbreak in humans in China in 2013.According to the latest data provided by the Chinese Center for Disease Control and Preven... Dear Editor,H7N9 is a recently identified subtype of influenza A virus that caused a major outbreak in humans in China in 2013.According to the latest data provided by the Chinese Center for Disease Control and Prevention(http://www.moh.gov.cn/zwgk/yqbb3/ejlist.shtml,updated on October 31,2018),the mortality rate of H7N9 infections in China amounts to 39.7%(611/1536).Thus,H7N9 poses a serious public health threat. 展开更多
关键词 identification NOVEL UNIVERSAL POTENTIAL epitope Cytoplasmic Tail
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新疆和田地区1株小鹅瘟病毒分离鉴定及VP基因的克隆分析
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作者 刘建华 孙文程 +4 位作者 胡月 吴桂灵 马彩红 撒瑞雪 阿力米古丽·乌米尔哈孜 《黑龙江畜牧兽医》 北大核心 2023年第15期84-89,共6页
为了找到新疆和田地区某鹅场雏鹅大量死亡的原因,试验采用PCR方法及琼脂扩散试验检测了病死鹅组织,将阳性病料经无菌处理后接种无免番鸭胚,盲传3代,进行病毒分离;设计GPV VP基因特异性引物对其尿囊液进行扩增,构建重组质粒和测序,分析V... 为了找到新疆和田地区某鹅场雏鹅大量死亡的原因,试验采用PCR方法及琼脂扩散试验检测了病死鹅组织,将阳性病料经无菌处理后接种无免番鸭胚,盲传3代,进行病毒分离;设计GPV VP基因特异性引物对其尿囊液进行扩增,构建重组质粒和测序,分析VP基因核苷酸序列同源性、亲缘关系,以及VP蛋白的抗原表位。结果表明:PCR检测及琼脂扩散试验结果均为阳性,其他常见病毒PCR检测结果为阴性;接种病料组织研磨悬液后无免番鸭胚出现典型症状,最终分离到一株GPV分离株,将其命名为GPV/GOOSE/XJHT/2020。该毒株VP基因核苷酸序列与GenBank中不同国家地区毒株的相似性均在95.0%以上;与强毒株DY16、RC16核苷酸序列的相似性均为99.3%,亲缘性高且处在同一分支;与疫苗株SYG61v、弱毒株GPV-98E的相似性均为95.0%,亲缘性较低且处在不同分支。VP蛋白具多个潜在的抗原表位。说明该鹅场存在GPV感染,且与国内毒株亲缘关系较近。 展开更多
关键词 小鹅瘟病毒 分离 鉴定 VP基因 进化分析 抗原表位预测
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非洲猪瘟病毒p54蛋白单克隆抗体制备及其抗原表位鉴定 被引量:7
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作者 齐艳丽 刘桃雪 +5 位作者 于海深 张超 鲁维飞 王江 褚贝贝 张改平 《畜牧兽医学报》 CAS CSCD 北大核心 2023年第1期281-292,共12页
旨在制备非洲猪瘟病毒(ASFV)p54蛋白的特异性单克隆抗体。本研究利用大肠杆菌表达系统表达p54蛋白,免疫BALB/c小鼠,取其脾细胞与SP2/0细胞进行细胞融合。利用纯化的p54蛋白作为包被抗原,采用间接ELISA方法筛选获得阳性杂交瘤细胞。经4... 旨在制备非洲猪瘟病毒(ASFV)p54蛋白的特异性单克隆抗体。本研究利用大肠杆菌表达系统表达p54蛋白,免疫BALB/c小鼠,取其脾细胞与SP2/0细胞进行细胞融合。利用纯化的p54蛋白作为包被抗原,采用间接ELISA方法筛选获得阳性杂交瘤细胞。经4次亚克隆后,取杂交瘤细胞上清进行单克隆抗体亚型鉴定,利用体内诱生法制备单克隆抗体并进行纯化。间接ELISA方法检测单克隆抗体的效价,利用交叉反应性试验、间接免疫荧光试验和蛋白印迹对所获单克隆抗体的特异性进行鉴定。根据预测的p54蛋白二级结构,采用逐步截短法分析鉴定单克隆抗体识别的抗原表位区域,并在p54的三级结构中进行标注。结果显示:成功筛选了6株分泌p54单克隆抗体的杂交瘤细胞,分别命名为28G12-1、31G7-1、31G7-2、35F10-1、35F10-2、38D3-1。其中28G12-1、31G7-1、31G7-2重链为IgG2a型,35F10-1、35F10-2、38D3-1重链为IgG1型;轻链均为κ链。单克隆抗体的最低效价为1∶25 600,与猪繁殖与呼吸综合征病毒、猪伪狂犬病病毒、猪流行性腹泻病毒、猪细小病毒、猪急性腹泻综合征冠状病毒、猪圆环病毒2型不发生交叉反应,特异性强,所获抗体均识别p54蛋白C端127—146 aa肽段。本研究成功获得了ASFV p54蛋白和p54单克隆抗体,并鉴定了6株单克隆抗体的抗原表位,为p54蛋白功能研究和ASFV新型表位疫苗研究奠定了基础。 展开更多
关键词 非洲猪瘟病毒 p54蛋白 原核表达 单克隆抗体 间接ELISA 抗原表位
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重组牛乳源金黄色葡萄球菌GapC蛋白优势B细胞抗原表位的预测和筛选
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作者 陈晓萌 张雪静 +2 位作者 张欢 张宝江 苏艳 《生物技术通报》 CAS CSCD 北大核心 2023年第5期306-313,共8页
旨在表达牛乳源金黄色葡萄球菌(Staphylococcus aureus)GapC蛋白并对其B细胞抗原表位进行预测与鉴定,本研究利用实验室分离鉴定的S.aureus分离株15119扩增GapC基因并构建重组表达质粒pET-28a-GapC,诱导纯化得到分子量为44 kD重组蛋白Ga... 旨在表达牛乳源金黄色葡萄球菌(Staphylococcus aureus)GapC蛋白并对其B细胞抗原表位进行预测与鉴定,本研究利用实验室分离鉴定的S.aureus分离株15119扩增GapC基因并构建重组表达质粒pET-28a-GapC,诱导纯化得到分子量为44 kD重组蛋白GapC,以此免疫新西兰大白兔,获得特异多克隆抗体。利用生物信息学方法,对GapC蛋白的二级及三级结构进行分析,预测其B细胞抗原表位,并利用特异性抗体对筛选的表位进行鉴定。结果表明,GapC蛋白具有良好的免疫原性,筛选出7个线性B细胞抗原表位,利用兔抗重组GapC蛋白多克隆抗体鉴定得到了PL 5(^(221)IPEIDGKLDGGAQRVP^(236))多肽和PL 7(^(264)KNASNESFGYTEDEIVSSDVVGM^(286))2个优势B细胞表位。本研究成功制备了GapC蛋白,预测并鉴定了2个优势抗原表位,为其嵌合表位疫苗的开发提供了技术支持。 展开更多
关键词 牛源金黄色葡萄球菌 GapC蛋白 表达 B细胞抗原表位 预测与鉴定
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无乳链球菌甘油醛-3-磷酸脱氢酶C的B细胞抗原表位预测与鉴定
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作者 李胜男 王汉青 +2 位作者 吕芬芬 赵海利 苏艳 《新疆农业大学学报》 CAS 2023年第4期277-284,共8页
旨在表达牛源无乳链球菌(Streptococcus agalactiae)甘油醛-3-磷酸脱氢酶C (GapC)蛋白并对其免疫效果进行评估,预测并鉴定其优势B细胞抗原表位。本研究对无乳链球菌X16087株的GapC基因扩增,构建重组质粒pET-28a-WR-X16087,经诱导表达后... 旨在表达牛源无乳链球菌(Streptococcus agalactiae)甘油醛-3-磷酸脱氢酶C (GapC)蛋白并对其免疫效果进行评估,预测并鉴定其优势B细胞抗原表位。本研究对无乳链球菌X16087株的GapC基因扩增,构建重组质粒pET-28a-WR-X16087,经诱导表达后纯化GapC蛋白,免疫小鼠并分析免疫效果。对GapC蛋白进行生物信息学分析,预测并鉴定其B细胞表位。结果表明,重组质粒经诱导表达纯化后,获得大小为44 kDa的目的蛋白,免疫小鼠后抗体效价可达1∶218 700,抗体亚型以IgG1和IgG2b为主;小鼠免疫后保护率可达100%;GapC免疫后可显著减少攻击后小鼠肝、脾、肺、肾中的菌体载量;GapC蛋白可减少细菌入侵对小鼠造成病理损伤。预测并筛选了GapC的B细胞表位,确定了3个具有良好抗原性的B细胞线性表位,分别为:~(46)KYDTTQGRFDGTVEVKDG~(63)、~(121)TAPGGNDV~(128)、~(186)MVLDGPHRGGDLRRAR~(201)。鉴定表明,3个B细胞表位有成为牛源无乳链球菌预防性表位疫苗的潜力。 展开更多
关键词 无乳链球菌 GapC蛋白 免疫效果 B细胞表位 筛选与鉴定
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抗原表位研究方法进展 被引量:30
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作者 宋帅 李春玲 +2 位作者 贾爱卿 杨冬霞 李淼 《动物医学进展》 CSCD 北大核心 2010年第12期87-91,共5页
抗原表位是抗原分子中的主要功能单位,能有效刺激机体的细胞免疫和体液免疫。随着免疫学和生物信息学技术的不断发展,用于研究T细胞表位和B细胞表位的方法得到了很大的提高。论文中概述了近几年用于研究T细胞表位的预测方法和鉴定方法,... 抗原表位是抗原分子中的主要功能单位,能有效刺激机体的细胞免疫和体液免疫。随着免疫学和生物信息学技术的不断发展,用于研究T细胞表位和B细胞表位的方法得到了很大的提高。论文中概述了近几年用于研究T细胞表位的预测方法和鉴定方法,以及在B细胞表位研究中所用的表位肽扫描技术、蛋白质"切割"法、噬菌体展示技术、X-衍射与核磁共振、表位预测方法等技术,并对每种研究方法进行了比较,为从事抗原表位研究的人员提供参考,从而更有利于表位肽疫苗的研制和诊断方法的建立。 展开更多
关键词 T细胞表位 B细胞表位 预测 鉴定
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肿瘤抗原MAGE-12的B细胞表位4分支多重抗原肽的合成及鉴定 被引量:10
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作者 李艳秋 吴玉章 +1 位作者 倪兵 贾正才 《第三军医大学学报》 CAS CSCD 北大核心 2002年第10期1143-1145,共3页
目的 设计合成肿瘤抗原MAGE 12的B细胞表位抗原肽并对其进行鉴定。方法 在预测MAGE 12的B细胞表位序列为QSDEGSSNEEQE(82 -93 )的基础上 ,采用 4分支多重抗原肽结构合成抗原肽 ,免疫C 5 7BL 6小鼠 ,产生多克隆抗体 ,应用ELISA实验及... 目的 设计合成肿瘤抗原MAGE 12的B细胞表位抗原肽并对其进行鉴定。方法 在预测MAGE 12的B细胞表位序列为QSDEGSSNEEQE(82 -93 )的基础上 ,采用 4分支多重抗原肽结构合成抗原肽 ,免疫C 5 7BL 6小鼠 ,产生多克隆抗体 ,应用ELISA实验及免疫组化实验检测多克隆抗体是否为人的抗原肽的特异性抗体。结果 免疫C 5 7BL 6小鼠后可产生MAGE 12的抗原特异性的多克隆抗体 ,抗体滴度可达 1∶64。结论 证明所预测的表位为MAGE 12的B细胞表位。 展开更多
关键词 肿瘤抗原 B细胞表位 MAGE-12 表位鉴定 酶联免疫吸附法 肿瘤免疫治疗
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抗原表位鉴定方法的研究进展 被引量:28
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作者 文雪霞 陈化兰 +1 位作者 熊永忠 王秀荣 《中国畜牧兽医》 CAS 北大核心 2012年第7期66-70,共5页
抗原表位是抗原分子中能与相应淋巴细胞表面的受体结合的特殊化学基团,一般由几个氨基酸残基序列或其空间结构组成。它是引起免疫应答的物质基础,其性质决定了抗原的特异性。抗原表位的鉴定方法主要有肽扫描技术、氨基酸定点突变技术、X... 抗原表位是抗原分子中能与相应淋巴细胞表面的受体结合的特殊化学基团,一般由几个氨基酸残基序列或其空间结构组成。它是引起免疫应答的物质基础,其性质决定了抗原的特异性。抗原表位的鉴定方法主要有肽扫描技术、氨基酸定点突变技术、X-ray衍射或核磁共振技术、质谱技术、噬菌体展示肽技术及生物信息技术预测等,文章将从这几方面入手对近几年来鉴定抗原表位的方法作一综述。 展开更多
关键词 抗原表位 鉴定方法 研究进展
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空肠弯曲菌主要外膜蛋白OMP18的B细胞抗原表位分析及功能鉴定 被引量:6
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作者 楼宏强 胡野 +3 位作者 王岚 单小云 盛秀胜 高素华 《中国人兽共患病学报》 CAS CSCD 北大核心 2014年第7期739-742,共4页
目的从生物信息学角度了解空肠弯曲菌主要外膜蛋白OMP18的跨膜结构、B细胞抗原表位及其抗原性、基因序列保守性等特征,并鉴定空肠弯曲菌主要外膜蛋白优势B细胞抗原表位,为后续抗体检测和疫苗研究提供实验依据。方法使用NCBI/Blast,TMHMM... 目的从生物信息学角度了解空肠弯曲菌主要外膜蛋白OMP18的跨膜结构、B细胞抗原表位及其抗原性、基因序列保守性等特征,并鉴定空肠弯曲菌主要外膜蛋白优势B细胞抗原表位,为后续抗体检测和疫苗研究提供实验依据。方法使用NCBI/Blast,TMHMM Server V2.0,DNA Star等生物信息学分析软件对空肠弯曲菌主要外膜蛋白OMP18进行蛋白跨膜结构预测、线性B细胞抗原表位及其抗原性分析、并对不同空肠弯曲菌OMP18蛋白的氨基酸序列同源性进行比较。采用空肠弯曲菌全菌抗体IgG为一抗,基于ELISA技术对优势线性B细胞抗原表位进行筛选鉴定。结果生物信息学预测结果表明外膜蛋白OMP18定位于空肠弯曲菌外膜且不存在跨膜结构。蛋白序列保守存在于各空肠弯曲菌株中,序列同源性达95%以上。同时,我们发现该外膜蛋白中存在3个明显的B细胞线性表位,且具有很强的抗原性。ELISA检测鉴定结果表明3个B细胞抗原表位都能有效识别空肠弯曲菌全菌抗体。结论空肠弯曲菌主要外膜蛋白OMP18保守存在于细菌表面,存在3个重要的B细胞线性抗原表位,可用于后续的抗体检测和疫苗开发研究。 展开更多
关键词 空肠弯曲菌 外膜蛋白OMP18 序列分析 表位鉴定 OMP18
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β-Netrin抗原表位的分析及其抗体的制备与鉴定 被引量:5
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作者 余利红 高艳 +3 位作者 王利红 韩洪彦 孙志贤 张成岗 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2005年第3期330-333,共4页
目的:制备抗神经细胞突起生长诱向因子(β-Netrin)的多克隆抗体(pAb)和单克隆抗体(mAb),并对其特异性进行鉴定。方法:应用GoldKey软件分析人βNetrinC末端结构域氨基酸的序列(共114个氨基酸),确定抗原表位并人工合成多肽。然后采用碳化... 目的:制备抗神经细胞突起生长诱向因子(β-Netrin)的多克隆抗体(pAb)和单克隆抗体(mAb),并对其特异性进行鉴定。方法:应用GoldKey软件分析人βNetrinC末端结构域氨基酸的序列(共114个氨基酸),确定抗原表位并人工合成多肽。然后采用碳化二亚胺法,将合成肽与载体蛋白牛血清白蛋白(BSA)偶联作为抗原,免疫BALB/c小鼠。取免疫小鼠脾细胞与Sp2/0骨髓瘤细胞常规融合,依次进行HAT选择培养,间接ELISA法筛选阳性的杂交瘤细胞并克隆化。对分泌的mAb的效价、Ig亚类(型)及特异性,分别用间接ELISA和Westernblot进行鉴定。同时,通过免疫细胞化学染色鉴定抗体的特异性。另外,以偶联的分子免疫新西兰白兔,制备抗β-Netrin的pAb,用Westernblot鉴定其特异性。结果:以确定的此16个氨基酸的序列NH2-FRGKRT-LYPESWTDRG-COOH作为抗原表位,以人工合成多肽与BSA偶联作为免疫原,获得3株可稳定分泌特异性抗β-Netrin mAb的杂交瘤细胞。免疫细胞化学染色结果表明,这3株抗体均能特异性地识别细胞中的抗原。另外,制备了高效价的抗β-Netrin的pAb,并能特异性地识别原核表达的β-Netrin蛋白。结论:采用人工合成多肽作为半抗原成功地制备出抗β-Netrin的pAb和mAb。 展开更多
关键词 神经细胞突起生长诱向因子 合成肽 抗原表位 抗体制备 抗体鉴定
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