Gutiérrez, José María

Karatt-­Vellatt, Aneesh

L. Olesen, Majken

Laustsen, Andreas Hougaard

Lomonte, Bruno

M. Luther, Alice

McCafferty, John

Slavny, Peter

W. Masters, Edward

Snakebite envenoming is a major cause of death and morbidity in tropical parts of the world. Current therapies are based on animal-­derived antisera that are associated with a high degree of immunogenicity, high cost, and batch-to-batch variation. Here, we report the results of our ongoing efforts of developing the world’s first fully recombinant antivenom based on human IgGs targeting the key toxins from the notorious black mamba (Dendroaspis polylepis)

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Downloaded from orbit.dtu.dk on: Dec 18, 2017Human oligoclonal recombinant antivenom against the black mamba (Dendroaspispolylepis)Laustsen, Andreas Hougaard; Karatt--Vellatt, Aneesh; Slavny, Peter; M. Luther, Alice; L. Olesen, Majken;W. Masters, Edward; Lomonte, Bruno; Gutiérrez, José María; McCafferty, JohnPublication date:2016Document VersionPublisher's PDF, also known as Version of recordLink back to DTU OrbitCitation (APA):Laustsen, A. H., Karatt--Vellatt, A., Slavny, P., M. Luther, A., L. Olesen, M., W. Masters, E., ... McCafferty, J.(2016). Human oligoclonal recombinant antivenom against the black mamba (Dendroaspis polylepis). Postersession presented at PEGS Europe: Protein & Antibody Engineering Summit 2016, Lisbon, Portugal.Human  oligoclonal recombinant  antivenom  against  the  black  mamba  (Dendroaspis polylepis)Andreas  H.  Laustsen1,2,  Aneesh Karatt-­‐‑Vellatt3;  Peter  Slavny3;  Alice  M.Luther3,  Majken L.  Olesen3Edward  W.  Masters3,  Bruno  Lomonte4;  José  María Gutiérrez4;  John  McCafferty31Department  of  Biotechnology  and  Biomedicine,  Technical  University  of  Denmark2Department  of  Drug  Design  and  Pharmacology,  Faculty  of  Health  and  Medical  Sciences,  University  of  Copenhagen3IONTAS  Ltd.,  Babraham Research  Institute,  Cambridge  CB22  3AT,  United  Kingdom  4Instituto  Clodomiro  Picado,  Facultad de  Microbiología,  Universidad  de  Costa  Rica,  San  José,  Costa  Rica    Contact informationDTU BioengeeringTechnical University of DenmarkSøltofts Plads 223DK-­‐‑2800 Kgs. Lyngby, Denmarkahola@bio.dtu.dk/ (+45) 2988 1134AcknowledgementsWe thank the Novo Nordisk Foundation (NNF16OC0019248), EliteForsk Rejsestipendie, Oticon Fonden,Lundbeckfonden, Fonden for Lægevidenskabens Fremme, Knud Højgaards Fond, Christian og OttiliaBrorsons rejselegat for yngre videnskabsmænd og kvinder,, IONTAS Ltd., and Instituto ClodomiroPicado for financial support.References[1]  Gutiérrez  JM,  León  G,  Lomonte  B,  Angulo  Y.  In-­‐‑flammation  &  Allergy-­‐‑Drug  Targets  2011,  10;  p369-­‐‑80.  [2]  Laustsen  AH,  Engmark  M,  Milbo  C,  Johannesen  J,  Lomonte  B,  Gutiérrez  JM,  Lohse  B.  Current  Pharmaceutical  Design  2016,  22.[3]  Laustsen  AH,  Lohse  B,  Lomonte  B,  Engmark  M,  Gutiérrez  JM.  Toxicon  2015,  104;  p43-­‐‑45.[4]  Laustsen  AH,  Lomonte  B,  Lohse  B,  Fernández  J,  Gutiérrez  JM.  Journal  of  Proteomics  2015,  119;  p126-­‐‑142.[5]  Laustsen  AH.  Recombinant  Antivenoms.  University  of  Copenhagen  2016.[6]  Laustsen  AH,  Johansen  KH,  Engmark  M,  Andersen  MR.  Nature  2016,  538;  p41.[7]  Laustsen  AH.  Toxin  Reviews  2016,  35;  p165-­‐‑170.scFvIgG scaffold+ +Human IgG1.  Panning4.  Elution5.  AmplificationRepeatAnalyze6.  ELISA2.  Binding 3.  WashingDiscovery of human scFvs by phage displayBased on a combined toxicovenomics and phage display selectionapproach5, 431 human scFv binders were isolated from a phagedisplay library against the medically relevant neurotoxins anddendrotoxins of D. polylepis.Conversion of unique scFv binders to IgGFollowing expression of 147 scFvs with unique VH CDR3 regions inE. coli and evaluation of binding strength, the 24 most promisingscFvs were selected and converted to IgG format. Of these, 20displayed good binding after successful expression in HEK cells.050000100000150000Emission,  615  nmn050000100000150000Emission,  615  nmn1 2 3 4 5 6 1 2 3 4 6 8 10 12 14 16 17 185 7 9 11 13 15⍺-­‐‑long  neurotoxin  IgGs ⍺-­‐‑dendrotoxin IgGsHuman oligoclonal recombinant antivenomsCurrently, the binding affinities and the neutralization potential ofthese 20 toxin-­‐‑targeting IgGs is being investigated with the aim ofbeing able to design a cost-­‐‑effective recombinant oligoclonalmixture of IgGs6 that can effectively neutralize the lethal effects ofthe synergistically-­‐‑acting venom of D. polylepis7.Short  neurotoxin Long  neurotoxin DendrotoxinSnakebite: a neglected tropical diseaseSnakebite envenoming is a major cause of death and morbidity intropical parts of the world. Current therapies are based on animal-­‐‑derived antisera that are associated with a high degree ofimmunogenicity, high cost, and batch-­‐‑to-­‐‑batch variation1,2. Here, wereport the results of our ongoing efforts of developing the world’sfirst fully recombinant antivenom based on human IgGs targetingthe key toxins3 from the notorious black mamba (Dendroaspispolylepis)4.Oligoclonal  expression  of  human  IgGs

Human oligoclonal recombinant antivenom against the black mamba (<i>Dendroaspis polylepis</i>)

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Human oligoclonal recombinant antivenom against the black mamba (<i>Dendroaspis polylepis</i>)

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