Functional single-cell hybridoma screening using droplet-based microfluidics

Bachir El Debs, Ramesh Utharala, Irina V. Balyasnikova, Andrew D. Griffiths, Christoph A. Merten*

*Corresponding author for this work

Research output: Contribution to journalArticle

126 Citations (Scopus)

Abstract

Monoclonal antibodies can specifically bind or even inhibit drug targets and have hence become the fastest growing class of human therapeutics. Although they can be screened for binding affinities at very high throughput using systems such as phage display, screening for functional properties (e.g., the inhibition of a drug target) is much more challenging. Typically these screens require the generation of immortalized hybridoma cells, as well as clonal expansion in microtiter plates over several weeks, and the number of clones that can be assayed is typically no more than a few thousand. We present here a microfluidic platform allowing the functional screening of up to 300,000 individual hybridoma cell clones within less than a day. This approach should also be applicable to nonimmortalized primary B-cells, as no cell proliferation is required: Individual cells are encapsulated into aqueous microdroplets and assayed directly for the release of antibodies inhibiting a drug target based on fluorescence. We used this system to perform a model screen for antibodies that inhibit angiotensin converting enzyme 1, a target for hypertension and congestive heart failure drugs. When cells expressing these antibodies were spiked into an unrelated hybridoma cell population in a ratio of 1:10,000 we observed a 9,400-fold enrichment after fluorescence activated droplet sorting. A wide variance in antibody expression levels at the single-cell level within a single hybridoma line was observed and high expressors could be successfully sorted and recultivated.

Original languageEnglish (US)
Pages (from-to)11570-11575
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume109
Issue number29
DOIs
StatePublished - Jul 17 2012

Fingerprint

Microfluidics
Hybridomas
Antibodies
Pharmaceutical Preparations
Clone Cells
Fluorescence
Peptidyl-Dipeptidase A
Bacteriophages
B-Lymphocytes
Heart Failure
Monoclonal Antibodies
Cell Proliferation
Hypertension
Population

Keywords

  • Angiotensin converting enzyme 1
  • Cell-based assay
  • High-throughput screening
  • Monoclonal antibody
  • Single-cell screening

ASJC Scopus subject areas

  • General

Cite this

Debs, Bachir El ; Utharala, Ramesh ; Balyasnikova, Irina V. ; Griffiths, Andrew D. ; Merten, Christoph A. / Functional single-cell hybridoma screening using droplet-based microfluidics. In: Proceedings of the National Academy of Sciences of the United States of America. 2012 ; Vol. 109, No. 29. pp. 11570-11575.
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Functional single-cell hybridoma screening using droplet-based microfluidics. / Debs, Bachir El; Utharala, Ramesh; Balyasnikova, Irina V.; Griffiths, Andrew D.; Merten, Christoph A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 109, No. 29, 17.07.2012, p. 11570-11575.

Research output: Contribution to journalArticle

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