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Detection of amino-terminal extracellular domain of somatostatin receptor 2 by specific monoclonal antibodies and quantification of receptor density in medulloblastoma.

dc.contributor.author Bigner, DD
dc.contributor.author Kuan, Chien-Tsun
dc.contributor.author Kumar, U
dc.contributor.author McLendon, Roger E
dc.contributor.author Wikstrand, CJ
dc.contributor.author Zalutsky, Michael Rod
dc.coverage.spatial United States
dc.date.accessioned 2011-04-15T16:46:29Z
dc.date.issued 2009-12
dc.identifier http://www.ncbi.nlm.nih.gov/pubmed/20025498
dc.identifier.uri http://hdl.handle.net/10161/3241
dc.description.abstract Somatostatin receptor 2 (SSTR2) is expressed by most medulloblastomas (MEDs). We isolated monoclonal antibodies (MAbs) to the 12-mer (33)QTEPYYDLTSNA(44), which resides in the extracellular domain of the SSTR2 amino terminus, screened the peptide-bound MAbs by fluorescence microassay on D341 and D283 MED cells, and demonstrated homogeneous cell-surface binding, indicating that all cells expressed cell surface-detectable epitopes. Five radiolabeled MAbs were tested for immunoreactive fraction (IRF), affinity (KA) (Scatchard analysis vs. D341 MED cells), and internalization by MED cells. One IgG(3) MAb exhibited a 50-100% IRF, but low KA. Four IgG(2a) MAbs had 46-94% IRFs and modest KAs versus intact cells (0.21-1.2 x 10(8) M(-1)). Following binding of radiolabeled MAbs to D341 MED at 4 degrees C, no significant internalization was observed, which is consistent with results obtained in the absence of ligand. However, all MAbs exhibited long-term association with the cells; binding at 37 degrees C after 2 h was 65-66%, and after 24 h, 52-64%. In tests with MAbs C10 and H5, the number of cell surface receptors per cell, estimated by Scatchard and quantitative FACS analyses, was 3.9 x 10(4) for the "glial" phenotype DAOY MED cell line and 0.6-8.8 x 10(5) for four neuronal phenotype MED cell lines. Our results indicate a potential immunotherapeutic application for these MAbs.
dc.language eng
dc.language.iso en_US
dc.relation.ispartof Hybridoma (Larchmt)
dc.relation.isversionof 10.1089/hyb.2009.0049
dc.subject Amino Acid Sequence
dc.subject Antibodies, Monoclonal
dc.subject Blotting, Western
dc.subject Cell Line, Tumor
dc.subject DNA Primers
dc.subject Enzyme-Linked Immunosorbent Assay
dc.subject Flow Cytometry
dc.subject Humans
dc.subject Immunohistochemistry
dc.subject Immunotherapy
dc.subject Medulloblastoma
dc.subject Protein Structure, Tertiary
dc.subject Receptors, Somatostatin
dc.subject Reverse Transcriptase Polymerase Chain Reaction
dc.title Detection of amino-terminal extracellular domain of somatostatin receptor 2 by specific monoclonal antibodies and quantification of receptor density in medulloblastoma.
dc.type Journal article
dc.description.version Version of Record
duke.date.pubdate 2009-12-0
duke.description.issue 6
duke.description.volume 28
dc.relation.journal Hybridoma
pubs.author-url http://www.ncbi.nlm.nih.gov/pubmed/20025498
pubs.begin-page 389
pubs.end-page 403
pubs.issue 6
pubs.organisational-group Clinical Science Departments
pubs.organisational-group Duke
pubs.organisational-group Duke Cancer Institute
pubs.organisational-group Institutes and Centers
pubs.organisational-group Neurosurgery
pubs.organisational-group Pathology
pubs.organisational-group Radiation Oncology
pubs.organisational-group Radiology
pubs.organisational-group School of Medicine
pubs.organisational-group Surgery
pubs.publication-status Published
pubs.volume 28
dc.identifier.eissn 1557-8348


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