Document Detail


Functional single-cell hybridoma screening using droplet-based microfluidics.
MedLine Citation:
PMID:  22753519     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Bachir El Debs; Ramesh Utharala; Irina V Balyasnikova; Andrew D Griffiths; Christoph A Merten
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Publication Detail:
Type:  Journal Article     Date:  2012-07-02
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  109     ISSN:  1091-6490     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2012 Jul 
Date Detail:
Created Date:  2012-07-18     Completed Date:  2012-10-11     Revised Date:  2013-07-12    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  11570-5     Citation Subset:  IM    
Affiliation:
Institut de Science et d'Ingénierie Supramoléculaires, Université de Strasbourg, 8 allée Gaspard Monge, 67083 Strasbourg Cedex, France.
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal / metabolism*
Antibody Affinity / physiology
B-Lymphocytes / metabolism
Fluorescence
High-Throughput Screening Assays / methods*
Hybridomas / metabolism*
Microfluidics / methods*
Peptidyl-Dipeptidase A / metabolism
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; EC 3.4.15.1/Peptidyl-Dipeptidase A
Comments/Corrections

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