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Identification and epitope mapping of anti-p72 single-chain antibody against African swine fever virus based on phage display antibody library

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农业科学学报:英文版 2023年第9期22卷 2834-2847页

作者： SONG Jin-xing WANG Meng-xiang ZHANG Yi-xuan WAN Bo DU Yong-kun ZHUANG Guo-qing LI Zi-bin QIAO Song-lin GENG Rui WU Ya-nan ZHANG Gai-ping College of Veterinary Medicine Henan Agricultural UniversityZhengzhou 450046P.R.China International Joint Research Center of National Animal Immunology Zhengzhou 450046P.R.China College of Animal Science and Veterinary Medicine Tianjin Agricultural UniversityTianjin 300384P.R.China Longhu Laboratory Zhengzhou 450046P.R.China School of Advanced Agricultural Sciences Peking UniversityBeijing 100871P.R.China Henan Provincial Key Laboratory of Animal Immunology Henan Academy of Agricultural SciencesZhengzhou 450002P.R.China

African swine fever virus(ASFV)is a lethal pathogen that causes severe threats to the global swine industry and it has already had catastrophic socio-economic effects.To date,no licensed prophylactic vaccine exists.Limited knowledge exists about the major immunogens of ASFV and the epitope mapping of the key antigens.As such,there is a considerable requirement to understand the functional monoclonal antibodies(mAbs)and the epitope mapping may be of utmost importance in our understanding of immune responses and designing improved vaccines,therapeutics,and diagnostics.In this study,we generated an ASFV antibody phage-display library from ASFV convalescent swine PBMCs,further screened a specific ASFV major capsid protein(p72)single-chain antibody and fused with an IgG Fc fragment(scFv-83-Fc),which is a specific recognition antibody against ASFV Pig/HLJ/2018 strain.Using the scFv-83-Fc mAb,we selected a conserved epitope peptide(221MTGYKH226)of p72 retrieved from a phage-displayed random peptide library.Moreover,flow cytometry and cell uptake experiments demonstrated that the epitope peptide can significantly promote BMDCs maturation in vitro and could be effectively uptaken by DCs,which indicated its potential application in vaccine and diagnostic reagent development.Overall,this study provided a valuable platform for identifying targets for ASFV vaccine development,as well as to facilitate the optimization design of subunit vaccine and diagnostic reagents.

关键词： ASFV phage display antibody library single chain antibody p72 epitope

Screening and identification of HLA-A2-restricted neoepitopes for immunotherapy of non-microsatellite instability-high colorectal cancer

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Science China(Life Sciences) 2022年第3期65卷 572-587页

作者： Ranran Shi Yubing Li Ling Ran Yu Dong Xiuman Zhou Jingwen Tang Lu Han Mingshuang Wang Liwei Pang Yuanming Qi Yahong Wu Yanfeng Gao School of Life Sciences Zhengzhou UniversityZhengzhou 450001China School of Pharmaceutical Sciences(Shenzhen) Sun Yat-sen UniversityShenzhen 518107China Henan Key Laboratory of Bioactive Macromolecules Zhengzhou UniversityZhengzhou 450001China Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital Zhengzhou 450008China Department of Integrated Chinse and Western Medicine Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer HospitalZhengzhou 450008China

Colorectal cancer has one of the highest mortality rates among malignant tumors,and most patients with non-microsatellite instability-high (MSI-H) colorectal cancer do not benefit from targeted therapy or immune checkpoint inhibitors.Identification of immunogenic neoantigens is a promising strategy for inducing specific antitumor T cells for cancer immunotherapy.Here,we screened potential high-frequency neoepitopes from non-MSI-H colorectal cancer and tested their abilities to induce tumorspecific cytotoxic T cell responses.Three HLA-A2-restricted neoepitopes (P31,P50,and P52) were immunogenic and could induce cytotoxic T lymphocytes in peripheral blood mononuclear cells from healthy donors and colorectal cancer patients.Cytotoxic T lymphocytes induced in HLA-A2.1/K^(b) transgenic mice could recognize and lyse mutant neoepitope-transfected HLA-A2^(+) cancer cells.Adoptive transfer of cytotoxic T lymphocytes induced by the peptide pool of these three neoepitopes effectively inhibited tumor growth and increased the therapeutic effects of anti-PD-1 antibody.These results revealed the potential of high-frequency mutation-specific peptide-based immunotherapy as a personalized treatment approach for patients with non-MSI-H colorectal cancer.The combination of adoptive T cell therapy based on these neoepitopes with immune checkpoint inhibitors,such as anti-PD-1,could provide a promising treatment strategy for non-MSI-H colorectal cancer.

关键词： non-MSI-H colorectal cancer PD-1 neoantigen epitope cancer immunotherapy

epitope Analysis of Anti-SARS-CoV-2 Neutralizing Antibodies

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Current Medical Science 2021年第6期41卷 1065-1074页

作者： Jun-biao XUE Sheng-ce TAO Shanghai Center for Systems Biomedicine Key Laboratory of Systems Biomedicine(Ministry of Education)Shanghai Jiao Tong UniversityShanghai 200240China

Coronavirus disease 2019 is threatening thousands of millions of people around the world.In the absence of specific and highly effective medicines,the treatment of infected persons is still very challenging.As therapeutics,neutralizing antibodies(NAbs)have great potential.Many NAbs have been reported,and most target various regions on the receptor-binding domain of the spike(S)protein,or the N-terminal domain.Several NAbs and NAb cocktails have been authorized for emergency use,and more arc in clinical trials or are under development.In this review,considering the angle of binding epitopes on the S protein,we summarize the functions and the underlying mechanisms of a set of well-recognized NAbs and provide guidance for vaccine design and the combinatorial use of these antibodies.In addition,we review the NAbs and NAb cocktails that have been approved for emergency use and discuss the effectiveness of these NAbs for combating severe acute respiratory syndrome coronavirus 2 mutants.

关键词： coronavirus disease 2019 severe acute respiratory syndrome coronavirus 2 neutralizing antibody epitope therapeutics vaccine

The amino acids differences in epitopes may promote the different allergenicity of ovomucoid derived from hen eggs and quail eggs

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Food Science and Human Wellness 2023年第3期12卷 861-870页

作者： Mengzhen Hao Shuai Yang Shiwen Han Huilian Che Key Laboratory of Precision Nutrition and Food Quality Key Laboratory of Functional DairyMinistry of EducationCollege of Food Science and Nutritional EngineeringChina Agricultural UniversityBeijing 100083China

Quail egg ovomucoid can inhibit activation of basophils and eosinophils,while hen egg ovomucoid has been shown to be a major allergen,named Gal d 1.At present,the differences in structure and function between two ovomucoid are unclear.We found the homology of ovomucoid in quail eggs and hen eggs reached77%.Compared with hen egg ovomucoid,the distribution of secondary structure was different in AA52-53,AA57-58,AA66-68,AA71-72,AA131-133,AA139-140,AA157-159 and AA184-185.Among 9 epitopes of egg ovomucoid,there were different amino acids from quail egg ovomucoid in 8 epitopes.Recombination quail egg ovomucoid had trypsin inhibition activity and quail egg ovomucoid didn't specifically bind to serum of eggs allergic patients.Quail egg ovomucoid can significantly inhibit RBL-2 H3 cells degranulation and protect cells morphology to a certain extent,indicating quail egg ovomucoid can inhibit cells activation and have potential anti-allergic effects,which is related to trypsin inhibitory activity.The difference in sensitization compare to hen egg ovomucoid may be due to amino acids differences affecting protein structure by changing antigenic epitopes.

关键词： Quail egg Hen egg Ovomucoid epitope Degranulation

Preparation and Initial Application of a Monoclonal Antibody Specific for a Newly Discovered Conserved Linear epitope of Rabies Virus Nucleoprotein

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Biomedical and Environmental Sciences 2012年第1期25卷 98-103页

作者： LV Xin Jun MA Xue Jun WANG Li Hua LI Hao SHEN Xin Xin YU Peng Cheng TANG Qing LIANG Guo Dong State Key Laboratory for Infectious Disease Prevention and Control Department of Viral Encephalitis Institute for Viral Disease Control and Prevention Chinese Centers for Disease Control and Prevention Beijing 102206 China State Key Laboratory for Molecular Virology and Genetic Engineering Department of Core Facility Institute for Viral Disease Control and Prevention Chinese Centers for Disease Control and Prevention Beijing 102206 China

Objective To prepare monoclonal antibodies against a newly discovered and conserved linear epitope of Rabies virus nucleoprotein and to use them in a rabies diagnostic test. Methods Synthetic peptide containing the epitope was used as immunogen to prepare hybridoma cell lines by classical hybridoma technology. Anti-peptide monoclonal antibodies produced in ascites of inoculated Balb/c mice were labeled with fluorescein isothiocyanate （FITC） after purification and used in fluorescent antibody test （FAT）. Results Two positive hybridoma cell lines, RVNP-mAbl-CL and RVNP-mAb2-CL, were obtained. RVNP- mAbl-CL produced a higher concentration of monoclonal antibody RVNP-mAbl in Balb/c ascites. FITC-labeled RVNP-mAbl showed correct results on certain Rabies virus-positive canine brain tissue samples and cells of a small subclone of baby hamster kidney 21 cell line （BSR）. Conclusion FITC-labeled RVNP-mAbl has potential application for laboratory diagnosis of rabies

关键词： Rabies virus Nucleoprotein epitope Peptide Monoclonal antibody

Rotavirus VP7 epitope chimeric proteins elicit cross-immunoreactivity in guinea pigs

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Virologica Sinica 2015年第5期30卷 363-370页

作者： Bingxin Zhao Xiaoxia Pan Yumei Teng Wenyue Xia Jing Wang Yuling Wen Yuanding Chen Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Diseases Institute of Medical BiologyChinese Academy of Medical Sciences & Peking Union Medical College Key Laboratory of Chemistry in Ethnic Medicinal Resources State Ethnic Affairs Commission and Ministry of EducationYunnan Minzu University

VP7 of group A rotavirus(RVA) contains major neutralizing epitopes. Using the antigenic protein VP6 as the vector, chimeric proteins carrying foreign epitopes have been shown to possess good immunoreactivity and immunogenicity. In the present study, using modified VP6 as the vector,three chimeric proteins carrying epitopes derived from VP7 of RVA were constructed. The results showed that the chimeric proteins reacted with anti-VP6 and with SA11 and Wa virus strains.Antibodies from guinea pigs inoculated with the chimeric proteins recognized VP6 and VP7 of RVA and protected mammalian cells from SA11 and Wa infection in vitro. The neutralizing activities of the antibodies against the chimeric proteins were significantly higher than those against the vector protein VP6 F. Thus, development of chimeric vaccines carrying VP7 epitopes using VP6 as a vector could be a promising alternative to enhance immunization against RVAs.

关键词： rotavirus VP7 epitope chimeric vaccine VP6 foreign

Identification of an epitope of SARS-coronavirus nucleocapsid protein

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Cell Research 2003年第3期13卷 141-145页

作者： YING LIN,XU SHEN,RUI FU YANG,YI XUE LI,YONG YONG JI,YOU Yu HE,MU DE SHI,WEI LU,TIE LIU SHI,JIN WANG,HONG XIA WANG,HUA LIANG JIANG,JIAN HUA SHEN,You HUA XIE,YUAN WANG,GANG PEI,BEI FEN SHEN,JIA RUI WU,BING SUNInstitute of Biochemistry and Cell Biology,Institute of Materia Medico,,Bioinformation Center,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences. Shanghai 200031,ChinaInstitute of Microbiology and Epidemiology,Institute of Basic Medical Sciences,Academy of Military Medical,Sc.imce.fi,Beijing 100071,China Institute of Biochemistry and Cell Biology Chinese Academy of Sciences Shanghai 200031 China.

The nucleocapsid(N)protein of severe acute respiratory syndrome-coronavirus(SARS-CoV)is a majorvirion structural protein.In this study,two epitopes(N1 and N2)of the N protein of SARS-CoV werepredicted by bioinformatics analysis.After immnnization with two peptides,the peptides-specific antibodieswere isolated from the immunized rabbits.The further experiments demonstrated that N1 peptide-inducedpolyclonal antibodies had a high affinity to bind to E.coli expressed Nprotein of SARS-CoV.Furthermore,itwas confirmed that N1 peptide-specific IgG antibodies were detectable in the sera of severe acute respiratorysyndrome(SARS)patients.The results indicated that an epitope of the N protein has been identified andN protein specific Abs were produced by peptide immunization,which will be useful for the study of SARS-CoV.

关键词： severe acute respiratory syndrome-coronavirus necleocapsid protein epitope polyclonal antibody antiserum.

In silico identification and characterization of common epitope-based peptide vaccine for Nipah and Hendra viruses

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Asian Pacific Journal of Tropical Medicine 2017年第6期10卷 601-608页

作者： Chayan Kumar Saha Mahbub Hasan Saddam Hossain Asraful Jahan Abul Kalam Azad Department of Genetic Engineering & Biotechnology Shahjalal University of Science and Technology Department of Genetic Engineering & Biotechnology University of Chittagong

Objective: To explore a common B-and T-cell epitope-based vaccine that can elicit an immune response against encephalitis causing genus Henipaviruses, Hendra virus(He V) and Nipah virus(Ni V). Methods: Membrane proteins F, G and M of He V and Ni V were retrieved from the protein database and subjected to different bioinformatics tools to predict antigenic B-cell epitopes. Best B-cell epitopes were then analyzed to predict their T-cell antigenic potentiality. Antigenic B-and T-cell epitopes that shared maximum identity with He V and Ni V were selected. Stability of the selected epitopes was predicted. Finally, the selected epitopes were subjected to molecular docking simulation with HLA-DR to confirm their antigenic potentiality in silico. Results: One epitope from G proteins, one from M proteins and none from F proteins were selected based on their antigenic potentiality. The epitope from the G proteins was stable whereas that from M was unstable. The M-epitope was made stable by adding flanking dipeptides. The 15-mer G-epitope(VDPLRVQWRNNSVIS) showed at least 66% identity with all Ni V and He V G protein sequences, while the 15-mer M-epitope(GKLEFRRNNAIAFKG) with the dipeptide flanking residues showed 73% identity with all Ni V and He V M protein sequences available in the database. Molecular docking simulation with most frequent MHC class-II(MHC II) and class-I(MHC I) molecules showed that these epitopes could bind within HLA binding grooves to elicit an immune response. Conclusions: Data in our present study revealed the notion that the epitopes from G and M proteins might be the target for peptide-based subunit vaccine design against He V and Ni V. However, the biochemical analysis is necessary to experimentally validate the interaction of epitopes individually with the MHC molecules through elucidation of immunity induction.

关键词： Hendra virus Nipah virus epitope Encephalitis Docking Flanking dipeptides

Identification and characterization of mammaglobin-A epitope in heterogenous breast cancers for enhancing tumor-targeting therapy

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Signal Transduction and Targeted Therapy 2020年第1期5卷 1778-1788页

作者： Zhiqiang Liu Xiqin Yang Cuimi Duan Jiangxue Li Rongsheng Tong Yuting Fan Jiannan Feng Ruiyuan Cao Wu Zhong Xiaoyan Feng Heqiu Zhang Lulu Cai Beijing Institute of Basic Medical Sciences 27 Taiping RoadHaidian DistrictBeijing 100850China Personalized Drug Therapy Key Laboratory of Sichuan Province Department of PharmacySichuan Provincial People’s HospitalUniversity of Electronic Science and Technology of ChinaChengdu 611731China Beijing Institute of Pharmacology and Toxicology 27 Taiping RoadHaidian DistrictBeijing 100850China

Although targeted therapy has been extensively investigated for breast cancers,a molecular target with broad application is currently unavailable due to the high heterogeneity of these cancers.Mammaglobin-A(Mam-A),which is overexpressed in most breast carcinomas,has been proposed as a promising target.However,the lack of specific targeting moieties due to uncertain binding epitopes hampers further translational study.Here,seven potential epitopes of Mam-A were disclosed,and a unique epitope was then identified in most types of breast cancers,despite the genotypic heterogeneity.With phage display technology,the epitope was determined to be N-terminal amino acids 42–51 of Mam-A(N42–51).Then,the N42–51 epitope-specific monoclonal antibody,mAb785,was conjugated to poly lactic-co-glycolic acid(PLGA)nanoparticles loaded with therapeutic agents,thereby enhancing the drug uptake and therapeutic efficacy in different genotypes of breast cancers.The computer simulation of the N42–51 epitope and the mAb785 structures,as well as their interactions,further revealed the specific targeting mechanism of the mAb785-conjugated nanoparticles to breast cancers.

关键词： breast cancers epitope