Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1
|
| pubmed:dateCreated |
1990-3-29
|
| pubmed:abstractText |
A procedure is described which permits high-yield direct cloning of newly established hybridomas on STO fibroblast feeders in soft agarose. Several thousand independent clones are typically obtained from each fusion (1 X 10(8) spleen cells). These are screened using a colony replica assay in which secreted antibodies diffuse through an agar overlay and bind to antigen immobilised on nitrocellulose. Bound antibodies are then detected with enzyme-labelled second antibody. The procedure is fast and efficient and permits the isolation and selection of antigen-specific clones in less than 2 weeks from fusion. It has been successfully employed for the derivation and selection of high-affinity anti-hapten antibodies. Other potential applications of the assay are in the detection of non-immobilised antigens by an indirect method using anti-globulin on nitrocellulose, in the generation of bispecific antibodies and the selection and characterisation of antibody specificities generated by the expression of antibody fragments in bacteria or yeasts.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Jan
|
| pubmed:issn |
0022-1759
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
24
|
| pubmed:volume |
126
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
61-8
|
| pubmed:dateRevised |
2003-11-14
|
| pubmed:meshHeading | |
| pubmed:year |
1990
|
| pubmed:articleTitle |
A single-step procedure for cloning and selection of antibody-secreting hybridomas.
|
| pubmed:affiliation |
Medical Research Council Laboratory of Molecular Biology, Cambridge, U.K.
|
| pubmed:publicationType |
Journal Article
|