Novel lymphocyte screening tube using dried monoclonal antibody reagents

Cytometry B Clin Cytom. Nov-Dec 2015;88(6):361-70. doi: 10.1002/cyto.b.21251. Epub 2015 Jul 17.

Abstract

We previously developed a 10-color 11-antibody combination including a viability dye, to screen T-, B-, and natural killer (NK)-cell populations in blood, bone marrow, tissue, and body fluids. Recently, Beckman Coulter has introduced a line of dried reagents that, unlike liquid reagents and cocktails, require no refrigeration, titration, or manipulation before using. We evaluated custom tubes based on our standard lymphocyte screening panel, focusing on comparative analysis, ease of use, and advantages compared with our liquid reagent set. We tested 42 samples from blood (n = 15), bone marrow (n = 17), and tissue (n = 10) with the combination CD4/CD8/KAPPA/LAMBDA/CD19/CD56/CD5/CD20/CD10/CD3/CD45 and a vital dye by both methods and compared positivity and staining intensity for each antigen. Of the 42 samples, 5 were normal samples, 3 were red cell disorders, 20 were B-cell malignancies, 5 T-cell malignancies, 4 myeloid malignancies, and the remaining 5 were other diagnoses. Dried reagents gave equivalent staining intensity results to our standard panel in a variety of sample types, with diagnoses including reactive lymphocytosis, chronic lymphocytic leukemia, and various lymphomas. Our standard panel for evaluation of mature lymphoid malignancies allows rapid assessment of any sample type while providing direct assessment of viability. The dried reagent tube reduces preanalytical work, with simple addition of sample and the viability dye to the tube, saving time, reducing potential errors, and obviating need to titrate and monitor individual antibodies. With a shelf life of at least 12 months, the reagents also offer potential savings in reagent costs by reducing wastage due to expiration or tandem breakdown in standard liquid formulation.

Keywords: acute leukemia; duraclone; flow cytometry; immunophenotyping; screening tube.

MeSH terms

  • Antibodies, Monoclonal / immunology*
  • Antigens, CD / immunology
  • Flow Cytometry / methods*
  • Humans
  • Immunophenotyping / methods*
  • Leukemia / immunology
  • Leukemia / pathology
  • Lymphocytes / immunology
  • Lymphocytes / pathology*
  • Lymphocytosis / diagnosis
  • Lymphocytosis / immunology
  • Lymphocytosis / pathology
  • Lymphoma / diagnosis
  • Lymphoma / immunology
  • Lymphoma / pathology

Substances

  • Antibodies, Monoclonal
  • Antigens, CD