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      Fractionation of normal and leukemic T-cells by lectin-affinity column chromatography

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      Cancer Letters
      Elsevier BV

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          Abstract

          A method for rapid fractionation of normal and leukemic T-cells (Jurkat, RPMI-8402, MOLT-4), using lectin-affinity column chromatography, is described. CNBr-activated Sepharose 6MB was used as a non-mobile phase. The gel was covalently conjugated with Dolichos biflorus agglutinin (DBA) over 24 h. The normal cells were eluted by phosphate buffered saline (Ca(2+) and Mg(2+) free), while the leukemic T-cells, interacting with DBA, were removed by N-acetyl-D-galactosamine or by low-concentrated acetic acid as a mobile phase. The cell fractions were detected spectrophotometrically at 600 nm. The rate of cell elution decreased in the order: normal>leukemic T-cells. The viability and the type of separated T-cell fractions were characterized by flow cytometry, using adequate fluorescent antibodies. The interactions between leukemic T-cells and DBA-saturated Sepharose beads were examined by fluorescent microscopy, using fluorescent isothiocyanate-DBA as a fluorescent marker.

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          Author and article information

          Journal
          Cancer Letters
          Cancer Letters
          Elsevier BV
          03043835
          October 2002
          October 2002
          : 184
          : 2
          : 207-214
          Article
          10.1016/S0304-3835(02)00207-0
          12127693
          5eb30796-6f25-48ef-a96d-9e07b22ce7d2
          © 2002

          https://www.elsevier.com/tdm/userlicense/1.0/

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