Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1-2
|
| pubmed:dateCreated |
1986-1-8
|
| pubmed:abstractText |
Spleen cells from non-immunized adult mice were fractionated on thin layers of fluorescein (FLU)-gelatin to yield FLU-specific B cells. These B cells were cultured either singly or in very small numbers in 10 microliter microcultures with 0.1 microgram/ml of the T cell-independent (TI) antigen FLU-E. coli lipopolysaccharide (FLU-LPS). Cultures were either filler cell-free, or supported by the addition of 10(5) CBA/N thymus cells per well. At 4-6 days, culture supernatants were assayed for the presence of anti-FLU antibody either by an enzyme-linked immunosorbent assay (ELISA) or a radioimmunoassay (RIA). With the filler cell-free cultures, B cell proliferation was scored microscopically before removal of culture supernatant. The cultured cells from each well were assayed for their capacity to form directly hemolytic FLU-specific plaques. In the filler cell-free system, the ELISA was much more sensitive than the plaque assay in identifying antibody-forming cell (AFC) clones, with over 10% of fractionated B cells yielding clones secreting detectable antibody, though with a low mean optical density (OD). This value represented over 80% of the proliferating clones. In the more efficient, filler cell-supported system, the difference between the 2 read-out methods was smaller. Here, one half of the hapten-specific B cells formed AFC clones, the highest cloning efficiency yet reported for an antigen-driven system. Comparative studies showed the RIA to be only marginally more sensitive than the ELISA, and not nearly as convenient for routine use.
|
| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies, Monoclonal,
http://linkedlifedata.com/resource/pubmed/chemical/Fluorescein,
http://linkedlifedata.com/resource/pubmed/chemical/Fluoresceins,
http://linkedlifedata.com/resource/pubmed/chemical/Gelatin,
http://linkedlifedata.com/resource/pubmed/chemical/Haptens
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Nov
|
| pubmed:issn |
0022-1759
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
28
|
| pubmed:volume |
84
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
327-41
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading |
pubmed-meshheading:3905971-Animals,
pubmed-meshheading:3905971-Antibodies, Monoclonal,
pubmed-meshheading:3905971-Antibody Affinity,
pubmed-meshheading:3905971-Antibody-Producing Cells,
pubmed-meshheading:3905971-B-Lymphocytes,
pubmed-meshheading:3905971-Cell Fractionation,
pubmed-meshheading:3905971-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:3905971-Female,
pubmed-meshheading:3905971-Fluorescein,
pubmed-meshheading:3905971-Fluoresceins,
pubmed-meshheading:3905971-Gelatin,
pubmed-meshheading:3905971-Haptens,
pubmed-meshheading:3905971-Hemolytic Plaque Technique,
pubmed-meshheading:3905971-Male,
pubmed-meshheading:3905971-Mice,
pubmed-meshheading:3905971-Radioimmunoassay,
pubmed-meshheading:3905971-Spleen,
pubmed-meshheading:3905971-Thymus Gland
|
| pubmed:year |
1985
|
| pubmed:articleTitle |
An ELISA assay efficiently detects clonal antibody formation by single, hapten-specific B lymphocytes.
|
| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
|