Microfluidic Separation of Lymphoblasts for the Isolation of Acute Lymphoblastic Leukemia Using the Human Transferrin Receptor as a Capture Target

Anal Chem. 2017 Jul 18;89(14):7340-7347. doi: 10.1021/acs.analchem.7b00377. Epub 2017 Jul 7.

Abstract

Acute lymphocytic leukemia (ALL) is the most prevalent pediatric cancer, and the peripheral blood lymphoblast percentage is an important index for ALL diagnosis and prognosis. We describe a microfluidic device that isolates and enumerates peripheral blood lymphoblasts using affinity separations. The innovative use of a nonspecific ligand allows a widespread "net" for cancer cells, without a priori knowledge of the cancer type. Using lymphoblasts spiked into blood, we simulated leukemia cases with lymphoblast concentrations ranging from 1 to 30% of total leukocytes. Lymphoblasts were isolated using monoclonal antibodies for the Human Transferring Receptor (CD71). Anti-CD71 antibodies were found to be more effective for capturing lymphoblasts than commonly used, ALL-specific antibodies for CD7 and CD10. CCRF-CEM lymphoblasts were isolated in the chip with 82-97% purity, with lower concentrations tested (7%) still showing >80% purity for cell capture. Patient-derived ALL cell lines COG-LL-332 and COG-LL-317 were isolated in the chip with 80%-97% and 57% -92% of purity, respectively, with the initial spike concentrations as low as 1%. The ability to capture ALL lymphoblasts present in blood at low concentrations provides a novel approach for characterization of ALL cells, including patients with low leukemic burdens during and after therapy.

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Antigen-Antibody Reactions
  • Antigens, CD / immunology*
  • Humans
  • Lymphocytes / immunology
  • Lymphocytes / pathology*
  • Microfluidic Analytical Techniques*
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / immunology
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / pathology*
  • Receptors, Transferrin / immunology*

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • CD71 antigen
  • Receptors, Transferrin