Characterization of a new monoclonal antibody against PAX5/BASP in 1525 paraffin-embedded human and animal tissue samples

Claudio Agostinelli, Elena Sabattini, Jakob Oemar Gjørret, Simona Righi, Maura Rossi, Manuela Mancini, Pier Paolo Piccaluga, Francesco Bacci, Teresa Marafioti, Giuliano Bettini, Brunangelo Falini, Stefano A. Pileri

Research output: Contribution to journalArticle

Abstract

We describe the newly generated DAK-PAX5 monoclonal antibody raised against a fixation-resistant epitope of the human PAX5/BSAP molecule. Materials and Methods: Following Western-blot, absorption, and chess-board titration tests, and optimization of antigen-retrieval and detection methods, DAK-Pax5 was used in parallel with a reference antibody (clone 24) on tissue micro-arrays (TMAs) constructed from normal human and animal tissues and from hematologic and nonhematologic human malignancies. Such TMAs were also tested with an anti-PAX2 antibody. Results: DAK-Pax5 reacted with normal human and animal B-cells and with 460/473 B-cell non-Hodgkin lymphomas (B-NHLs). All plasmacytomas/ plasmablastic tumors (n=13) and T/NK-cell neoplasms (n=264) turned out consistently negative as did acute myelogenous leukaemias (n=19) except 2 carrying t(8;21). Positivity was found in 6/6 and 155/169 lymphocyte predominant and classical HLs, respectively, although the staining intensity varied through cases. Among 521 nonhematologic malignancies, DAK-Pax5 reacted with 22/399 carcinomas (4/11 neuroendocrine, 2/4 Merkel-cell, 4/21 prostatic, 1/11 urothelial, 1/26 renal, 2/12 cervical squamous-cell, 3/13 ovarian, and 5/75 colonic). When compared with clone 24, DAK-Pax5 produced a stronger positivity in most if not all B-NHLs and HLs. No cross-reactivity with the anti-PAX2 antibody was recorded. Discussion: DAK-Pax5 represents a new reliable tool for diagnostics and research.

Original languageEnglish
Pages (from-to)561-572
Number of pages12
JournalApplied Immunohistochemistry and Molecular Morphology
Volume18
Issue number6
DOIs
Publication statusPublished - Dec 2010

Keywords

  • immunohistochemistry
  • normal tissues
  • PAX2
  • PAX5
  • tissue microarray
  • tumors

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Medical Laboratory Technology
  • Histology

Fingerprint Dive into the research topics of 'Characterization of a new monoclonal antibody against PAX5/BASP in 1525 paraffin-embedded human and animal tissue samples'. Together they form a unique fingerprint.

  • Cite this

    Agostinelli, C., Sabattini, E., Gjørret, J. O., Righi, S., Rossi, M., Mancini, M., Piccaluga, P. P., Bacci, F., Marafioti, T., Bettini, G., Falini, B., & Pileri, S. A. (2010). Characterization of a new monoclonal antibody against PAX5/BASP in 1525 paraffin-embedded human and animal tissue samples. Applied Immunohistochemistry and Molecular Morphology, 18(6), 561-572. https://doi.org/10.1097/PAI.0b013e3181e79013