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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:dateCreated |
1994-6-27
|
| pubmed:abstractText |
Western blot analysis proved that all cluster-2 MAbs recognize identical or overlapping disulfide-bond-dependent epitopes, indicating the presence of a disulfide-bond-stabilized EGP-2 domain carrying highly immunodominant non-linear epitopes. The apparent immunodominance of this domain makes it difficult to generate and select antibodies against other potentially useful EGP-2 epitopes. Using PCR, we have generated mutant EGP-2 cDNA (delta EGP-2) from which the coding sequences for a putative immunodominant 6-kDa intra-chain loop structure has been removed. delta EGP-2 transfected COS-7 cells reacted with MM104, an antibody detecting a linear epitope on EGP-2, but were not recognized by any cluster-2 MAb. To generate new anti-EGP-2 antibodies we constructed another mutant EGP-2 protein (delta EGP-2) from which additional domains, irrelevant for antibody generation (signal peptide, trans-membrane and cytoplasmic domains), were removed. delta EGP-2 was introduced in a prokaryotic expression system that adds a polyhistidine affinity tag to the delta EGP-2 N-terminus, making possible one-step purification by immobilized metal-ion-affinity chromatography (IMAC). Western blot analysis showed that sera derived from mice immunized with purified delta EGP-2 had high-titer antibodies to reduced EGP-2 samples. We conclude that the availability of prokaryotic and eukaryotic EGP-2-expression constructs might facilitate the selection of new anti-EGP-2 MAbs otherwise difficult to obtain.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies, Monoclonal,
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, Neoplasm,
http://linkedlifedata.com/resource/pubmed/chemical/Cell Adhesion Molecules,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Complementary,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Primers,
http://linkedlifedata.com/resource/pubmed/chemical/Epitopes,
http://linkedlifedata.com/resource/pubmed/chemical/Tumor Markers, Biological,
http://linkedlifedata.com/resource/pubmed/chemical/tumor-associated antigen GA733
|
| pubmed:status |
MEDLINE
|
| pubmed:issn |
0898-6924
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
8
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
64-9
|
| pubmed:dateRevised |
2007-7-23
|
| pubmed:meshHeading |
pubmed-meshheading:7515032-Amino Acid Sequence,
pubmed-meshheading:7515032-Animals,
pubmed-meshheading:7515032-Antibodies, Monoclonal,
pubmed-meshheading:7515032-Antigens, Neoplasm,
pubmed-meshheading:7515032-Base Sequence,
pubmed-meshheading:7515032-Blotting, Western,
pubmed-meshheading:7515032-Carcinoma, Small Cell,
pubmed-meshheading:7515032-Cell Adhesion Molecules,
pubmed-meshheading:7515032-Cell Line,
pubmed-meshheading:7515032-Cercopithecus aethiops,
pubmed-meshheading:7515032-DNA, Complementary,
pubmed-meshheading:7515032-DNA Primers,
pubmed-meshheading:7515032-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:7515032-Epitopes,
pubmed-meshheading:7515032-Humans,
pubmed-meshheading:7515032-Lung Neoplasms,
pubmed-meshheading:7515032-Molecular Sequence Data,
pubmed-meshheading:7515032-Polymerase Chain Reaction,
pubmed-meshheading:7515032-Restriction Mapping,
pubmed-meshheading:7515032-Transfection,
pubmed-meshheading:7515032-Tumor Cells, Cultured,
pubmed-meshheading:7515032-Tumor Markers, Biological
|
| pubmed:year |
1994
|
| pubmed:articleTitle |
Epitope mapping of SCLC-cluster-2 MAbs and generation of antibodies directed against new EGP-2 epitopes.
|
| pubmed:affiliation |
Department of Clinical Immunology, University Hospital Groningen, The Netherlands.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|