Refined protocol for generating monoclonal antibodies from single human and murine B cells

Details

• Alternative Title:
  J Immunol Methods
• Personal Author:
  Ho, Irvin Y. ; Bunker, Jeffrey J. ; Erickson, Steven A. ; Neu, Karlynn E. ; Huang, Min ; Cortese, Mario ; Pulendran, Bali ; Wilson, Patrick C.
• Description:
  Generating monoclonal antibodies from single B cells is a valuable tool for characterizing the specificity and functional properties of humoral responses. We and others developed protocols that have facilitated major advances in our understanding of B cell development, tolerance, and effector responses to HIV and influenza. Here, we demonstrate various refinements and dramatically reduce the time required to produce recombinant antibodies. Further, we present new methods for cloning and isolating antibodies from cells with lower immunoglobulin mRNA levels that may be resistant to traditional techniques. Together, these refinements significantly increase single-cell antibody expression efficiency and are easily integrated into established and novel pipelines.
• Subjects:
  Animals Antibodies, Monoclonal Antibody Expression Antibody Specificity Article B-Lymphocytes Cloning, Molecular Genetic Vectors Gibson Assembly Human Antibodies Humans Mice Monoclonal Antibodies Recombinant Proteins Single-cell PCR

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• Source:
  J Immunol Methods. 438:67-70
• Pubmed ID:
  27600311
• Pubmed Central ID:
  PMC5322767
• Document Type:
  Journal Article
• Funding:
  R37 AI048638/NIAID NIH HHS/National Institute of Allergy and Infectious Diseases Extramural Activities/United States ; F30 AI124476/NIAID NIH HHS/National Institute of Allergy and Infectious Diseases Extramural Activities/United States ; U19 AI090023/NIAID NIH HHS/National Institute of Allergy and Infectious Diseases Extramural Activities/United States ; P30 DK042086/NIDDK NIH HHS/National Institute of Diabetes and Digestive and Kidney Diseases/United States ; P51 OD011132/ODCDC CDC HHS/Office of the Director/United States ; U19 AI057266/NIAID NIH HHS/National Institute of Allergy and Infectious Diseases Extramural Activities/United States ; HHSN272201400006C/NIAID NIH HHS/National Institute of Allergy and Infectious Diseases Extramural Activities/United States ; T32 GM007281/NIGMS NIH HHS/National Institute of General Medical Sciences/United States ; U19 AI109946/NIAID NIH HHS/National Institute of Allergy and Infectious Diseases Extramural Activities/United States ; P01 AI097092/NIAID NIH HHS/National Institute of Allergy and Infectious Diseases Extramural Activities/United States ; U19 AI082724/NIAID NIH HHS/National Institute of Allergy and Infectious Diseases Extramural Activities/United States
• Volume:
  438
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:83b4d7da2f163744c59ba362399332de8d064f2aa0dcacd1b298960c391b10ff
• Download URL:
  https://stacks.cdc.gov/view/cdc/80482/cdc_80482_DS1.pdf
• File Type:
  [PDF - 1.12 MB ]