title: Evaluation of somatostatin receptor subtype expression in human neuroendocrine tumors using two sets of new monoclonal antibodies.
abstract: BACKGROUND
  The expression and reliable detection of somatostatin receptor subtypes (SSTR1-5) is a prerequisite for the successful use of somatostatin analogs in neuroendocrine tumors (NETs). Two sets of monoclonal antibodies (mAbs) against human SSTR1, 2A, 3 and 5 have recently been developed by two independent laboratories using rabbit and mouse hybridomas. Our aim was to evaluate the usefulness of both sets of mAbs for detection of SSTRs in NET samples as they are routinely collected in clinical practice.
  
  
  METHODS
  Mouse and rabbit mAbs were characterized in SSTR1, 2A, 3 and 5-transfected HEK293 cells and human archival samples of pancreatic tissue and NET. Comparative analysis of mAbs was also conducted by immunostaining of a tissue microarray composed of 75 cores of NET.
  
  
  RESULTS
  Immunohistochemical analysis of HEK293 cells showed that both rabbit and mouse mAbs specifically detect their cognate receptor subtype, with mild cytoplasmic cross-reactivity observed for rabbit mAbs. Both sets of mAbs labeled normal pancreatic islets and showed similar patterns of immunoreactivity in NET controls. Direct comparison of mAb sets using a NET tissue microarray revealed strong correlation between rabbit and mouse mAbs against SSTR1 and 5, and moderate correlation for SSTR3. The rabbit mAb against SSTR2A showed higher affinity for its cognate receptor than the corresponding mouse mAb, resulting in a more reliable detection of this SSTR.
  
  
  CONCLUSIONS
  mAbs from both sets are reliable tools for the detection of SSTR1, 3 and 5, whereas the rabbit mAb against SSTR2A is recommended for use in routine clinical testing due to its superior binding affinity.
authors.affiliation: Novartis Pharma AG, CH-4057 Basel, Switzerland. Electronic address: chiara.lambertini@novartis.com.
authors.forename: Chiara
authors.initials: C
authors.lastname: Lambertini
authors.forename: Patrizia
authors.initials: P
authors.lastname: Barzaghi-Rinaudo
authors.forename: Lisa
authors.initials: L
authors.lastname: D'Amato
authors.forename: Stefan
authors.initials: S
authors.lastname: Schulz
authors.forename: Paolo
authors.initials: P
authors.lastname: Nuciforo
authors.forename: Herbert A
authors.initials: HA
authors.lastname: Schmid
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry*; D000918:Antibody Specificity; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D005770:Gastrointestinal Neoplasms / Q000378:metabolism* / Q000473:pathology; D057809:HEK293 Cells; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D018358:Neuroendocrine Tumors / Q000378:metabolism* / Q000473:pathology; D010190:Pancreatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D016612:Paraffin Embedding; D011817:Rabbits; D017481:Receptors, Somatostatin / Q000276:immunology / Q000378:metabolism*; D046888:Tissue Array Analysis; D016707:Tissue Fixation
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D005404:Fixatives; D017481:Receptors, Somatostatin; D005557:Formaldehyde
keywords: Antibody; FFPE; GEP; IHC; NETs; Neuroendocrine tumors; Pituitary; SSTR; Somatostatin; Somatostatin analog; Somatostatin receptor; TMA; formalin-fixed paraffin-embedded; gastroenteropancreatic; immunohistochemical; mAb; monoclonal antibody; neuroendocrine tumors; reverse transcription polymerase chain reaction; rtPCR; somatostatin receptor subtype; tissue microarray