title: Carb-3 is the superior anti-CD15 monoclonal antibody for immunohistochemistry.
abstract: Immunohistochemical detection of CD15 is important in the diagnosis of Hodgkin lymphoma and may play a role in the classification of renal cell tumors (RCTs). In the NordiQC external quality assessment scheme, 4 CD15 tests, each with 71 to 121 participating laboratories, showed that 24% to 50% of the stains were insufficient. This was mainly because of very low primary antibody (Ab) concentration and insufficient heat-induced epitope retrieval, whereas the Ab clone performance seemed of little importance. The purpose of this study was to evaluate the performance of the most commonly used CD15 Abs on the basis of vendor-recommended and in-house optimized protocols. Multitissue blocks with 199 specimens including various malignant lymphomas, RCTs, and normal tissues were stained with 3 different concentrated (conc) CD15 Ab clones Carb-3, MMA, and BY87 according to predetermined in-house optimized protocols on 2 automated immunostaining platforms. Carb-3 and MMA were also applied in ready-to-use (RTU) formats utilized according to vendor protocols. Extension and intensity of stains was determined using the H-score method. Clone Carb-3-conc gave with an in-house optimized protocol the highest H-scores in Hodgkin lymphoma, RCTs, and normal kidney tissue. Clones Carb-3-RTU and MMA-conc gave slightly lower scores, whereas clones MMA-RTU and BY87-conc gave the lowest scores and a large proportion of false-negative reactions. For all concentrated Abs, in-house optimized protocols resulted in increased sensitivity and improved overall staining results compared with vendor-recommended protocols. The importance of Ab selection and protocol optimization in immunohistochemical laboratories is emphasized.
authors.affiliation: Institute of Pathology, Aalborg Hospital, Aarhus University Hospital.
authors.forename: Rasmus
authors.initials: R
authors.lastname: Røge
authors.forename: Søren
authors.initials: S
authors.lastname: Nielsen
authors.forename: Mogens
authors.initials: M
authors.lastname: Vyberg
mesh_terms: D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry* / Q000276:immunology; D000912:Antibodies, Neoplasm / Q000737:chemistry* / Q000276:immunology; D005260:Female; D005647:Fucosyltransferases / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D016256:Lewis X Antigen / Q000276:immunology / Q000378:metabolism*; D008297:Male; D009369:Neoplasms / Q000276:immunology / Q000378:metabolism* / Q000473:pathology*
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D000912:Antibodies, Neoplasm; D016256:Lewis X Antigen; C066693:FUT4 protein, human; D005647:Fucosyltransferases