About: A Rapid Method to Characterize Mouse IgG Antibodies and Isolate Native Antigen Binding IgG B Cell Hybridomas

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About: A Rapid Method to Characterize Mouse IgG Antibodies and Isolate Native Antigen Binding IgG B Cell Hybridomas Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	A Rapid Method to Characterize Mouse IgG Antibodies and Isolate Native Antigen Binding IgG B Cell Hybridomas
Creator	Jiang, Chengyu Ju, Xiangwu Liu, Kangtai Zhang, Gongyi Crawford, Frances Jin, Niyun Liu, Haolin Marrack, Philippa White, Janice Kappler, John
Source	Medline; PMC
abstract	B cell hybridomas are an important source of monoclonal antibodies. In this paper, we developed a high-throughput method to characterize mouse IgG antibodies using surface plasmon resonance technology. This assay rapidly determines their sub-isotypes, whether they bind native antigen and their approximate affinities for the antigen using only 50 μl of hybridoma cell culture supernatant. Moreover, we found that mouse hybridomas secreting IgG antibodies also have membrane form IgG expression without Igα. Based on this surface IgG, we used flow cytometry to isolate rare γ2a isotype switched variants from a γ2b antibody secreting hybridoma cell line. Also, we used fluorescent antigen to single cell sort antigen binding hybridoma cells from bulk mixture of fused hybridoma cells instead of the traditional multi-microwell plate screening and limiting dilution sub-cloning thus saving time and labor. The IgG monoclonal antibodies specific for the native antigen identified with these methods are suitable for in vivo therapeutic uses, but also for sandwich ELISA assays, histology, flow cytometry, immune precipitation and x-ray crystallography.
has issue date	2015-08-28(xsd:dateTime)
bibo:doi	10.1371/journal.pone.0136613
bibo:pmid	26317987
has license	cc-by
sha1sum (hex)	7266d5a2b588f4bd08e301590247156d59a7f227
schema:url	https://doi.org/10.1371/journal.pone.0136613
resource representing a document's title	A Rapid Method to Characterize Mouse IgG Antibodies and Isolate Native Antigen Binding IgG B Cell Hybridomas
has PubMed Central identifier	PMC4552657
has PubMed identifier	26317987
schema:publication	PLoS One
resource representing a document's body	covid:7266d5a2b588f4bd08e301590247156d59a7f227#body_text
is schema:about of	named entity 'native' named entity 'cell culture' named entity 'isotype' named entity 'hybridoma' named entity 'secreting' named entity 'mouse' named entity 'Igα' named entity 'resonance' named entity 'surface' named entity 'sort' named entity 'monoclonal antibodies' named entity 'hybridomas' named entity 'HYBRIDOMAS' named entity 'IGG B CELL' named entity 'CELL SURFACE' named entity 'HYBRIDOMA CELL LINE' named entity 'X-RAY CRYSTALLOGRAPHY' named entity 'ANTIBODY' named entity 'SUB-' named entity 'BCL' named entity 'IGG MONOCLONAL' named entity 'ASSAY' named entity 'TRADITIONAL' named entity 'THESE' named entity 'BIACORE' named entity 'RAPIDLY' named entity 'CLONING' named entity 'FIG' named entity 'RARE' named entity 'SECRETING' named entity 'MICROWELL PLATE' named entity 'SOURCE' named entity 'ANTIBODIES' named entity 'IGG' named entity 'SURFACE PLASMON RESONANCE' named entity 'TESTING' named entity 'ANTIBODIES SPECIFIC' named entity 'IN VIVO' named entity 'MEMBRANE FORM' named entity 'TO CHARACTERIZE' named entity 'IMMUNE PRECIPITATION' named entity 'USED' named entity 'FLUORESCENT' named entity 'SCREENING' named entity 'ANTIGEN' named entity 'CELL CULTURE SUPERNATANT' named entity 'NATIVE' named entity 'EXPRESSION' named entity 'HISTOLOGY' named entity 'STAINING' named entity 'THROUGHPUT' named entity 'ENRICHMENT' named entity 'USING' named entity 'VARIANTS' named entity 'METHOD' named entity 'IDENTIFIED' named entity 'HAVE' named entity 'MIXTURE' named entity 'SURFACE IGG' named entity 'ELISPOT' named entity 'HYBRIDOMAS' named entity 'CELL SORTING' named entity 'MACS' named entity 'ISOLATE' named entity 'PAPER' named entity 'HIGH' named entity '28A' named entity 'THEIR' named entity 'SWITCHED' named entity 'MONOCLONAL ANTIBODIES' named entity 'BULK' named entity 'BASED'

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