期刊
JOURNAL OF IMMUNOLOGICAL METHODS
卷 438, 期 -, 页码 67-70出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2016.09.001
关键词
Monoclonal antibodies; Single-cell PCR; Human antibodies; Gibson assembly; Antibody expression
资金
- National Institute of Allergy and Infectious Diseases
- National Institutes of Health [U19AI109946, P01AI097092, U19AI057266, U19AI05766-11, U19AI082724, U19AI090023]
- Department of Health and Human Services (CEIRS) [HHSN272201400006C]
- NIH grant [F30AI124476]
- NIH Medical Scientist Training Program grant [T32GM007281]
- University of Chicago Digestive Diseases Research Core Center [P30DK42086]
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. (C) 2016 Elsevier B.V. All rights reserved.
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