Facile Discovery of a Diverse Panel of Anti-Ebola Virus Antibodies by Immune Repertoire Mining.
The ongoing evolution of Ebolaviruses poses significant challenges to the development of immunodiagnostics for detecting emergent viral variants. There is a critical need for the discovery of monoclonal antibodies with distinct affinities and specificities for different Ebolaviruses. We developed an efficient technology for the rapid discovery of a plethora of antigen-specific monoclonal antibodies from immunized animals by mining the VH:VL paired antibody repertoire encoded by highly expanded B cells in the draining popliteal lymph node (PLN). This approach requires neither screening nor selection for antigen-binding. Specifically we show that mouse immunization with Ebola VLPs gives rise to a highly polarized antibody repertoire in CD138(+) antibody-secreting cells within the PLN. All highly expanded antibody clones (7/7 distinct clones/animal) were expressed recombinantly, and shown to recognize the VLPs used for immunization. Using this approach we obtained diverse panels of antibodies including: (i) antibodies with high affinity towards GP; (ii) antibodies which bound Ebola VLP Kissidougou-C15, the strain circulating in the recent West African outbreak; (iii) non-GP binding antibodies that recognize wild type Sudan or Bundibugyo viruses that have 39% and 37% sequence divergence from Ebola virus, respectively and (iv) antibodies to the Reston virus GP for which no antibodies have been reported.
Department of Chemical Engineering, University of Texas at Austin, Austin, Texas, USA.
Bo
B
Wang
Department of Molecular Biosciences, University of Texas at Austin, Austin, Texas, USA.
Christien A
CA
Kluwe
Department of Chemical Engineering, University of Texas at Austin, Austin, Texas, USA.
Oana I
OI
Lungu
Department of Chemical Engineering, University of Texas at Austin, Austin, Texas, USA.
Brandon J
BJ
DeKosky
Department of Molecular Biosciences, University of Texas at Austin, Austin, Texas, USA.
Scott A
SA
Kerr
Department of Chemical Engineering, University of Texas at Austin, Austin, Texas, USA.
Erik L
EL
Johnson
Hidetaka
H
Tanno
Chang-Han
Lee
Department of Chemical Engineering, University of Texas at Austin, Austin, Texas, USA.
Jiwon
J
Jung
Department of Chemical Engineering, University of Texas at Austin, Austin, Texas, USA.
Alec B
AB
Rezigh
Department of Chemical Engineering, University of Texas at Austin, Austin, Texas, USA.
Sean M
SM
Carroll
Department of Virology and Immunology, Texas Biomedical Research Institute, San Antonio, TX, USA.
Ann N
AN
Reyes
Department of Virology and Immunology, Texas Biomedical Research Institute, San Antonio, TX, USA.
Janelle R
JR
Bentz
Department of Biodefense and Protein Diagnostics, Luminex Corporation, Austin, TX, USA.
Itamar
I
Villanueva
Department of Biodefense and Protein Diagnostics, Luminex Corporation, Austin, TX, USA.
Amy L
AL
Altman
Department of Virology and Immunology, Texas Biomedical Research Institute, San Antonio, TX, USA.
Robert A
RA
Davey
Center for Systems and Synthetic Biology University of Texas at Austin, Austin, TX, USA.
Andrew D
AD
Ellington
Department of Chemical Engineering, University of Texas at Austin, Austin, Texas, USA.|Department of Molecular Biosciences, University of Texas at Austin, Austin, Texas, USA.|Center for Systems and Synthetic Biology University of Texas at Austin, Austin, TX, USA.|Institute for Cell and Molecular Biology, University of Texas at Austin, Austin, TX, USA.|Department of Biomedical Engineering, University of Texas at Austin, Austin, Texas, USA.
George
G
Georgiou
D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology*; D000917:Antibody Formation / Q000235:genetics / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D003429:Cross Reactions; D004195:Disease Models, Animal; D029043:Ebolavirus / Q000276:immunology*; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D019142:Hemorrhagic Fever, Ebola / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007114:Immunization; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D008198:Lymph Nodes / Q000276:immunology; D051379:Mice; D010641:Phenotype; D011485:Protein Binding / Q000276:immunology
D000914:Antibodies, Viral; D000939:Epitopes; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region