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

TitleRefined protocol for generating monoclonal antibodies from single human and murine B cells.
Publication TypeJournal Article
Year of Publication2016
AuthorsHo IY, Bunker JJ, Erickson SA, Neu KE, Huang M, Cortese M, Pulendran B, Wilson PC
JournalJ Immunol Methods
Volume438
Pagination67-70
Date Published2016 Nov
ISSN1872-7905
Abstract

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.

DOI10.1016/j.jim.2016.09.001
Alternate JournalJ. Immunol. Methods
PubMed ID27600311
PubMed Central IDPMC5322767
Grant ListR37 AI048638 / AI / NIAID NIH HHS / United States
F30 AI124476 / AI / NIAID NIH HHS / United States
U19 AI090023 / AI / NIAID NIH HHS / United States
P30 DK042086 / DK / NIDDK NIH HHS / United States
U19 AI057266 / AI / NIAID NIH HHS / United States
HHSN272201400006C / AI / NIAID NIH HHS / United States
T32 GM007281 / GM / NIGMS NIH HHS / United States
U19 AI109946 / AI / NIAID NIH HHS / United States
P01 AI097092 / AI / NIAID NIH HHS / United States
U19 AI082724 / AI / NIAID NIH HHS / United States
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