Microfluidic Separation of Lymphoblasts for the Isolation of Acute Lymphoblastic Leukemia Using the Human Transferrin...

Area of Science:

• Biomedical Engineering
• Oncology
• Hematology

Background:

• Acute lymphocytic leukemia (ALL) is the most common pediatric cancer.
• Peripheral blood lymphoblast percentage is crucial for ALL diagnosis and prognosis.
• Current diagnostic methods may require improvement for early detection and monitoring.

Purpose of the Study:

• To develop and validate a microfluidic device for isolating and enumerating peripheral blood lymphoblasts.
• To evaluate the efficacy of a nonspecific ligand and anti-CD71 antibodies for lymphoblast capture.
• To assess the device's performance in simulated and patient-derived leukemia samples.

Main Methods:

• A microfluidic device utilizing affinity separations was designed.
• Lymphoblasts were isolated using monoclonal antibodies targeting the Human Transferring Receptor (CD71).
• The device was tested with lymphoblasts spiked into blood at varying concentrations (1-30%) and with patient-derived ALL cell lines.

Main Results:

• Anti-CD71 antibodies demonstrated superior lymphoblast capture compared to ALL-specific markers (CD7, CD10).
• High purity (82-97%) of CCRF-CEM lymphoblasts was achieved, even at low concentrations (7%).
• Patient-derived ALL cell lines (COG-LL-332, COG-LL-317) were isolated with 80-97% and 57-92% purity, respectively, at concentrations as low as 1%.

Conclusions:

• The developed microfluidic device offers a novel and effective approach for capturing ALL lymphoblasts from peripheral blood.
• This technology enables accurate characterization of ALL cells, particularly in patients with low leukemic burdens.
• The device holds potential for improved ALL diagnosis, prognosis, and therapeutic monitoring.