Multiplexed screening of natural humoral immunity identifies antibodies at fine specificity for complex and dynamic viral targets

MAbs. Mar-Apr 2014;6(2):460-73. doi: 10.4161/mabs.27760. Epub 2014 Jan 8.

Abstract

Viral entry targets with therapeutic neutralizing potential are subject to multiple escape mechanisms, including antigenic drift, immune dominance of functionally irrelevant epitopes, and subtle variations in host cell mechanisms. A surprising finding of recent years is that potent neutralizing antibodies to viral epitopes independent of strain exist, but are poorly represented across the diverse human population. Identifying these antibodies and understanding the biology mediating the specific immune response is thus difficult. An effective strategy for meeting this challenge is to incorporate multiplexed antigen screening into a high throughput survey of the memory B cell repertoire from immune individuals. We used this approach to discover suites of cross-clade antibodies directed to conformational epitopes in the stalk region of the influenza A hemagglutinin (HA) protein and to select high-affinity anti-peptide antibodies to the glycoprotein B (gB) of human cytomegalovirus. In each case, our screens revealed a restricted VH and VL germline usage, including published and previously unidentified gene families. The in vivo evolution of paratope specificity with optimal neutralizing activity was understandable after correlating biological activities with kinetic binding and epitope recognition. Iterative feedback between antigen probe design based on structure and function information with high throughput multiplexed screening demonstrated a generally applicable strategy for efficient identification of safe, native, finely tuned antibodies with the potential for high genetic barriers to viral escape.

Keywords: broadly protective antibodies; cytomegalovirus; fusion; human antibodies; immunoglobulin germline; influenza; monoclonal antibodies; neutralizing antibodies; viral epitopes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Blocking / immunology
  • Antibodies, Blocking / metabolism*
  • Antibody Affinity
  • Antigens, Viral / immunology
  • Antigens, Viral / metabolism*
  • B-Lymphocyte Subsets / immunology*
  • B-Lymphocytes / immunology*
  • Cell Line
  • Cytomegalovirus / immunology*
  • Cytomegalovirus Infections / immunology*
  • Cytomegalovirus Infections / therapy
  • Epitopes / metabolism*
  • Hemagglutinin Glycoproteins, Influenza Virus / immunology
  • Hemagglutinin Glycoproteins, Influenza Virus / metabolism*
  • High-Throughput Screening Assays
  • Humans
  • Immune Evasion / drug effects
  • Immunity, Humoral
  • Immunity, Innate
  • Immunologic Memory
  • Influenza A virus / immunology*
  • Influenza, Human / immunology*
  • Influenza, Human / therapy
  • Protein Conformation
  • Viral Envelope Proteins / immunology
  • Viral Envelope Proteins / metabolism*

Substances

  • Antibodies, Blocking
  • Antigens, Viral
  • Epitopes
  • Hemagglutinin Glycoproteins, Influenza Virus
  • Viral Envelope Proteins
  • glycoprotein B, Simplexvirus