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

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.

AFM-compatible microfluidic platform for affinity-based capture and nanomechanical characterization of circulating tumor cells

“…The reversible physical bonding easily peels off of the PDMS chip for further AFM analysis of the captured intact CTCs residing on the glass substrate. While HB-based microfluidic devices have been widely used to isolate CTCs from blood samples of patients with epithelial (such as prostate 12 and breast 13 ) as well as nonepithelial (such as bone marrow 14 and lymphocytic leukemia 15 ) cancers, reversible microfluidic device assembly-which allows direct measurements of AFM tip-cell surface interactions with no restrictions on the external access of captured CTCs-to the best of our knowledge is novel.…”

Tandem microfluidic chip isolation of prostate and breast cancer cells from simulated liquid biopsies using CD71 as an affinity ligand

“…33 Mammography screening and MRI screening are used to detect tumors at the early stages. 34 We propose the use of anti-CD71 in combination with a high throughput 5-channel herringbone chip design which has a detection limit of two orders of magnitude lower than we have previously reported for standard herringbone devices, 23,24 and an order of magnitude lower than nanoparticle coated herringbone chips. 35 This chip can detect PC-3 and MDA-MB-231 cells at a 0.1% spike in lysed blood, and has several advantages over a single channel herringbone chip.…”

“…Estrogen receptors (ER) and progesterone receptors (PR), and HER2 biomarkers are used for the detection and treatment of breast cancer. 24,27 Triple negative breast cancers, which account for 10-20% of breast cancers, do not express ER, PR or HER2 receptors. [28][29][30] Prostate cancer is the 5 th leading cause of death in men, worldwide.…”

Continuous Separation of Circulating Tumor Cells from Whole Blood Using a Slanted Weir Microfluidic Device

“…The results imply that the antibody-based positive enrichment of CTCs may lose distinct sub-populations that possess crucial information regarding metastasis, and lead to a biased conclusion [2]. In order to overcome the limitation, other antibodies that are specific to a certain cancer types [12,13,14,15], or the use of a cocktail of tumor specific antibodies, have been reported [16]. However, these methods require information regarding the specific target cancer type or mutation beforehand, or require costly imaging equipment and reagents to visualize the antibody cocktail.…”