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Global characterization of modifications to the charge isomers of IgG antibody

Journal of Pharmaceutical Analysis 2022年第1期12卷 156-163页

作者： Xinling Cui Wei Mi Zhishang Hu Xiaoyu Li Bo Meng Xinyuan Zhao Xiaohong Qian Tao Zhu Wantao Ying State Key Laboratory of Proteomics Beijing Proteome Research CenterNational Center for Protein Sciences(Beijing)Beijing Institute of LifeomicsBeijingChina National Institute of Metrology Center for Advanced Measurement ScienceBeijingChina National Institute of Metrology Chemical Metrology&Analytical Science DivisionBeijingChina CanSino Bio Corporation Tianjin Key Laboratory of Recombinant Bacterial Recombination and Bacterial Vaccines TianjinChina

Posttranslational modifications of antibody products affect their stability,charge distribution,and drug activity and are thus a critical quality attribute.The comprehensive mapping of antibody modifications and different charge isomers(CIs)is of utmost importance,but is challenging.We intended to quantitatively characterize the posttranslational modification status of CIs of antibody drugs and explore the impact of posttranslational modifications on charge heterogeneity.The CIs of antibodies were fractionated by strong cation exchange chromatography and verified by capillary isoelectric focusing-whole column imaging detection,followed by stepwise structural characterization at three levels.First,the differences between CIs were explored at the intact protein level using a top-down mass spectrometry approach;this showed differences in glycoforms and deamidation status.Second,at the peptide level,common modifications of oxidation,deamidation,and glycosylation were identified.Peptide mapping showed nonuniform deamidation and glycoform distribution among CIs.In total,10 N-glycoforms were detected by peptide mapping.Finally,an in-depth analysis of glycan variants of CIs was performed through the detection of enriched glycopeptides.Qualitative and quantitative analyses demonstrated the dynamics of 24 N-glycoforms.The results revealed that sialic acid modification is a critical factor accounting for charge heterogeneity,which is otherwise missed in peptide mapping and intact molecular weight analyses.This study demonstrated the importance of the comprehensive analyses of antibody CIs and provides a reference method for the quality control of biopharmaceutical analysis.

关键词： antibody Charge isomers Mass spectrometry Posttranslational modification Glycopeptide

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Structural and functional analysis of a potent human neutralizing antibody against enterovirus A71

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Science China(Life Sciences) 2022年第12期65卷 2517-2526页

作者： Zhe Chen Linlin Bao Bin Zhu Hua Fu Shuangli Zhu Tianjiao Ji Ying Xue Chuan Liu Xurong Wang Fengdi Li Qi Lv Feifei Qi Pin Yu Wei Deng Wenbo Xu Chuan Qin Hongrong Liu Qi Jin MOH Key Laboratory of Systems Biology of Pathogens Institute of Pathogen BiologyChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100730China Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases NHC Key Laboratory of Human Disease Comparative MedicineInstitute of Laboratory Animal ScienceChinese Academy of Medical Sciences and Comparative Medicine CenterPeking Union Medical CollegeBeijing 100021China School of Computer Science and Engineering Hunan University of Science and TechnologyXiangtan 411201China National Institute for Viral Disease Control and Prevention China CDCBeijing 102206China Ministry of Education Key Laboratory of Protein Sciences Beijing Advanced Innovation Center for Structural BiologyTsinghua UniversityBeijing 100084China Key Laboratory for Matter Microstructure and Function of Hunan Province Key Laboratory of Low-dimensional Quantum Structures and Quantum ControlSchool of Physics and ElectronicsHunan Normal UniversityChangsha 410081China Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases Hangzhou 310058China

Enterovirus A71(EV-A71) causes major outbreaks of hand,foot,and mouth disease(HFMD) in many countries,most frequently affecting children,and a small proportion of cases may lead to death.Currently,no vaccine is available in most endemic regions,and no licenced treatments for EV-A71 infection are available.Here,we characterize a human monoclonal antibody(Hu MAb),E1,by screening a Fab antibody phage library derived from patients who recovered from EV-A71 infection.E1 exhibits strong neutralizing activity against EV-A71 virus in cells.The cryo-electron microscopy(cryo-EM) structures of the EV-A71 virion in complex with E1 Fab fragments demonstrated that E1 recognized an epitope formed by residues in the BC and HI loops of VP1.In a mouse model,E1 effectively protected against lethal EV-A71 challenge in both prophylactic and therapeutic treatment.In particular,E1 significantly reduces virus titers and muscle damage.E1 might represent a potential adjunct to EV-A71 treatment.

关键词： enterovirus A71 antibody mouse models cryo-electron microscopy

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antibody responses in COVID-19 patients

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The Journal of Biomedical Research 2020年第6期34卷 410-415页

作者： Shuying Liu Shan Lu SL Consulting Thousand OaksCA 91320USA Department of Medicine University of Massachusetts Medical SchoolWorcesterMA 01605USA.

Measuring virus-specific antibody responses to emerging pathogens is a well-established and highly useful tool to diagnose such infections,understand interactions between the immune system and pathogens,and provide potential clues for the development of vaccines or therapeutic agents against such pathogens.Since the beginning of 2020,the discovery of SARS-CoV-2 as the emerging virus responsible for the COVID-19 pandemic has provided new insight into the complexity of antibody responses to this dangerous virus.The current review aims to sort out diverse and sometimes seemingly confusing findings to put together a cohesive understanding on the profile of antibody responses elicited in COVID-19 patients.

关键词： COVID-19 antibody serology SARS-CoV-2

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Application of Oral Fluid in Measles IgG antibody Detection for Medical Staff

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Biomedical and Environmental Sciences 2022年第1期35卷 64-68页

作者： DONG Mei SUN Jing Yi XIE Hui LIU Jie WANG Yi Ting FU Yin DONG Jian Ping HUANG Fang Institute for immunization and prevention Beijing Centre for Disease Prevention and ControlBeijing 100013China Department of Infectious Diseases Beijing Haidian Hospital(Beijing Haidian Section of Peking University Third Hospital)Beijing 100080China Chaoyang District Centre for Disease Prevention and Control Beijing 100021China

Measles is an acute,highly contagious,and severe respiratory disease caused by the measles virus.Infected patients can develop fever,maculopapular rashes,pneumonia,encephalitis,and other complications,including death;.A measles epidemic has persisted in China for years.Crosscontamination can easily occur in hospitals,and medical staff is at high risk for cross-infection.

关键词： Measles antibody acute

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Molecular Insights into Striking antibody Evasion of SARS-CoV-2 Omicron Variant

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Chinese Physics Letters 2022年第10期39卷 71-78页

作者：阎增帅徐耀丁泓铭马余强 National Laboratory of Solid State Microstructures and Department of Physics Collaborative Innovation Center of Advanced MicrostructuresNanjing UniversityNanjing 210093China Center for Soft Condensed Matter Physics and Interdisciplinary Research School of Physical Science and TechnologySoochow UniversitySuzhou 215006China

The SARS-CoV-2 Omicron variant has become the dominant variant in the world.Uncovering the structural basis of altered immune response and enhanced transmission of Omicron is particularly important.Here,taking twenty-five antibodies from four groups as examples,we comprehensively reveal the underlying mechanism of how mutations in Omicron induces the weak neutralization by using molecular simulations.Overall,the binding strength of 68%antibodies is weakened in Omicron,much larger than that in Delta(40%).Specifically,the percentage of the weakened antibodies vary largely in different groups.Moreover,the mutation-induced repulsion is mainly responsive for the weak neutralization in AB/CD groups but does not take effect in EF group.Significantly,we demonstrate that the disappearance of hydrophobic interaction and salt bridges due to residue deletions contributes to the decreased binding energy in NTD group.This work provides unprecedented atomistic details for the distinct neutralization of WT/Delta/Omicron,which informs prospective efforts to design antibodies/vaccines against Omicron.

关键词： antibody details bridges

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antibody drug conjugate:the“biological missile”for targeted cancer therapy

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Signal Transduction and Targeted Therapy 2022年第4期7卷 1066-1090页

作者： Zhiwen Fu Shijun Li Sifei Han Chen Shi Yu Zhang Department of Pharmacy Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022People’s Republic of China Hubei Province Clinical Research Center for Precision Medicine for Critical Illness Wuhan 430022People’s Republic of China Drug Delivery Disposition and DynamicsMonash Institute of Pharmaceutical SciencesMonash University(Parkville Campus)381 Royal ParadeParkvilleVIC 3052Australia Faculty of Pharmacy China Pharmaceutical University639 Longmian AvenueJiangning DistrictNanjing 211198People’s Republic of China

antibody–drug conjugate(ADC)is typically composed of a monoclonal antibody(mAbs)covalently attached to a cytotoxic drug via a chemical linker.It combines both the advantages of highly specific targeting ability and highly potent killing effect to achieve accurate and efficient elimination of cancer cells,which has become one of the hotspots for the research and development of anticancer drugs.Since the first ADC,Mylotarg®(gemtuzumab ozogamicin),was approved in 2000 by the US Food and Drug Administration(FDA),there have been 14 ADCs received market approval so far worldwide.Moreover,over 100 ADC candidates have been investigated in clinical stages at present.This kind of new anti-cancer drugs,known as“biological missiles”,is leading a new era of targeted cancer therapy.Herein,we conducted a review of the history and general mechanism of action of ADCs,and then briefly discussed the molecular aspects of key components of ADCs and the mechanisms by which these key factors influence the activities of ADCs.Moreover,we also reviewed the approved ADCs and other promising candidates in phase-3 clinical trials and discuss the current challenges and future perspectives for the development of next generations,which provide insights for the research and development of novel cancer therapeutics using ADCs.

关键词： antibody drugs conjugate

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An Immunocompetent Patient with High Neutralizing antibody Titers Who Shed COVID-19 Virus for 169 days——China,2020

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China CDC weekly 2021年第32期3卷 688-691,I0003-I0008页

作者： Li Li Siqi Li Yang Pan Ling Qin Siyuan Yang Dawei Tan Yaling Hu Maria Deloria Knoll Xiaoli Wang Linghang Wang Quanyi Wang Beijing Ditan Hospital Capital Medical UniversityBeijingChina Johns Hopkins Bloomberg School of Public Health BaltimoreMarylandUSA Beijing Center for Disease Control and Prevention BeijingChina Biomedical Information Center of Beijing YouAn Hospital Capital Medical UniversityBeijingChina Sinovac Biotech Co LtdBeijingChina

Few reports are available about prolonged shedding of coronavirus disease(COVID-19)virus,also known as severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),by immunocompetent patients.We report a case of an infected,immunocompetent person with 169 days of COVID-19 virus shedding during which time he had two brief periods when he tested negative for COVID-19 followed by again testing positive.We describe relevant clinical,immunological,and genomic features.We found that continuous and prolonged viral replication and infectivity existed in an immunocompetent COVID-19 patient,despite having high neutralizing antibody titers.

关键词： antibody acute respiratory

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Thrombosis after SARS-CoV2 infection or COVID-19 vaccination:will a nonpathologic anti-PF4 antibody be a solution?—A narrative review

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Journal of Bio-X Research 2022年第3期5卷 97-103页

作者： Elizabeth Rao Payal Grover Hongtao Zhang Department of Pathology and Laboratory Medicine Perelman School of MedicineUniversity of PennsylvaniaPhiladelphiaPAUSA

The coronavirus disease 2019(COVID-19)pandemic was triggered by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),a previously unknown strain of coronavirus.To fully understand the consequences and complications of SARS-CoV-2 infections,we have reviewed current literature on coagulation dysfunctions that are related to the disease and vaccination.While COVID-19 is more commonly considered as a respiratory illness,studies indicate that,in addition to respiratory illness,a coagulation dysfunction may develop in individuals after the initial infection,placing them at the risk of developing thrombotic events.Patients who died of COVID-19 had higher levels of D-dimer,a biomarker for blood clot formation and breakdown.Effective treatments for coagulation dysfunctions are critically needed to improve patient survival.On the other hand,antibodies against platelet factor 4(PF4)/heparin may be found in patients with rare instances of vaccine-induced immunological thrombotic thrombocytopenia(VITT)following vaccination with adenovirus-based vaccines.VITT is characterized by atypical thrombosis and thrombocytopenia,similar to immune-mediated heparin-induced thrombocytopenia(HIT),but with no need for heparin to trigger the immune response.Although both adenovirus-based and mRNA-based vaccines express the Spike protein of SARS-CoV-2,VITT is exclusively related to adenovirus-based vaccines.Due to the resemblance with HIT,the use of heparin is highly discouraged against treating patients with thrombotic thrombocytopenia after SARS-CoV-2 infection or with VITT after vaccination.Intravenous immunoglobulin therapy coupled with anticoagulation is recommended instead.The well-studied anti-PF4 monoclonal antibody RTO,which does not induce pathologic immune complexes in the presence of heparin and has been humanized for a potential treatment modality for HIT,may provide a nonanticoagulant HIT-specific solution to the problem of increased blood coagulation after SARS-CoV-2 infection or the VITT after immunization.

关键词： antibody coagulation dysfunctions COVID-19 PF4 vaccine-induced immunological thrombotic thrombocytopenia

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Effects of CXCR7-neutralizing antibody on neurogenesis in the hippocampal dentate gyrus and cognitive function in the chronic phase of cerebral ischemia

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Neural Regeneration Research 2020年第6期15卷 1079-1085页

作者： Bing-Chao Dong Mei-Xuan Li Xiao-Yin Wang Xi Cheng Yu Wang Ting Xiao Jukka Jolkkonen Chuan-Sheng Zhao Shan-Shan Zhao Department of Neurology The First Hospital of China Medical UniversityShenyangLiaoning ProvinceChina Key Laboratory of Immunodermatology Ministry of HealthMinistry of EducationShenyangLiaoning ProvinceChina Department of Neurology Institute of Clinical MedicineUniversity of Eastern FinlandKuopioFinland

Stromal cell-derived factor-1 and its receptor CXCR4 are essential regulators of the neurogenesis that occurs in the adult hippocampal dentate gyrus.However,the effects of CXCR7,a new atypical receptor of stromal cell-derived factor-1,on hippocampal neurogenesis after a stroke remain largely unknown.Our study is the first to investigate the effect of a CXCR7-neutralizing antibody on neurogenesis in the dentate gyrus and the associated recovery of cognitive function of rats in the chronic stage of cerebral ischemia.The rats were randomly divided into sham,sham+anti-CXCR7,ischemia and ischemia+anti-CXCR7 groups.Endothelin-1 was injected in the ipsilateral motor cortex and striatum to induce focal cerebral ischemia.Sham group rats were injected with saline instead of endothelin-1 via intracranial injection.Both sham and ischemic rats were treated with intraventricular infusions of CXCR7-neutralizing antibodies for 6 days 1 week after surgery.Immunofluorescence staining with doublecortin,a marker for neuronal precursors,was performed to assess the neurogenesis in the dentate gyrus.We found that anti-CXCR7 antibody infusion enhanced the proliferation and dendritic development of doublecortin-labeled cells in the dentate gyrus in both ischemic and sham-operated rats.Spatial learning and memory functions were assessed by Morris water maze tests 30-32 days after ischemia.CXCR7-neutralizing antibody treatment significantly reduced the escape latency of the spatial navigation trial and increased the time spent in the target quadrant of spatial probe trial in animals that received ischemic insult,but not in sham operated rats.These results suggest that CXCR7-neutralizing antibody enhances the neurogenesis in the dentate gyrus and improves the cognitive function after cerebral ischemia in rats.All animal experimental protocols and procedures were approved by the Institutional Animal Care and Use Committee of China Medical University(CMU16089 R)on December 8,2016.

关键词： cerebral ischemia cognitive function CXCR7 dendritic development dentate gyrus doublecortin neurogenesis neutralizing antibody stroke stromal cell-derived factor-1

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Recent advances of antibody drug conjugates for clinical applications

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Acta Pharmaceutica Sinica B 2020年第9期10卷 1589-1600页

作者： Pengxuan Zhao Yuebao Zhang Wenqing Li Christopher Jeanty Guangya Xiang Yizhou Dong Division of Pharmaceutics&Pharmacology College of Pharmacythe Ohio State UniversityColumbusOH 43210USA School of Pharmacy Tongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China Department of Biomedical Engineering the Ohio State UniversityColumbusOH 43210USA The Center for Clinical and Translational Science the Ohio State UniversityColumbusOH 43210USA The Comprehensive Cancer Center the Ohio State UniversityColumbusOH 43210USA Dorothy M.Davis Heart&Lung Research Institute the Ohio State UniversityColumbusOH 43210USA Department of Radiation Oncology the Ohio State UniversityColumbusOH 43210USA Neuroscience Program College of Medicine&College of Arts and Sciencesthe Ohio State UniversityColumbusOH 43210USA

antibody drug conjugates(ADCs)normally compose of a humanized antibody and small molecular drug via a chemical linker.After decades of preclinical and clinical studies,a series of ADCs have been widely used for treating specific tumor types in the clinic such as brentuximab vedotin(Adcetris?)for relapsed Hodgkin’s lymphoma and systemic anaplastic large cell lymphoma,gemtuzumab ozogamicin(Mylotarg?)for acute myeloid leukemia,ado-trastuzumab emtansine(Kadcyla?)for HER2-positive metastatic breast cancer,inotuzumab ozogamicin(Besponsa?)and most recently polatuzumab vedotin-piiq(Polivy?)for B cell malignancies.More than eighty ADCs have been investigated in different clinical stages from approximately six hundred clinical trials to date.This review summarizes the key elements of ADCs and highlights recent advances of ADCs,as well as important lessons learned from clinical data,and future directions.

关键词： antibody drug conjugates antibody Cytotoxic agents Linker Clinical application

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