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

Abstract

Introduction: 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.