Isolation of monocytes from human peripheral blood using immuno-affinity expanded-bed adsorption.

Area of Science:

• Biotechnology
• Immunology
• Cell Biology

Background:

• Efficient separation of monocytes from human peripheral blood is crucial for research and clinical applications.
• Existing cell separation techniques can be time-consuming and may affect cell viability.

Purpose of the Study:

• To develop and characterize a novel technique for isolating monocytes using expanded-bed adsorption.
• To evaluate the efficiency, purity, and viability of monocytes separated by this new method.

Main Methods:

• Utilized expanded-bed adsorption with perfluorocarbon supports derivatized with avidin/streptavidin.
• Employed biotinylated monoclonal antibodies for indirect positive or negative cell capture.
• Investigated contactor design and operating conditions for selective cell isolation from peripheral blood mononuclear cells (PBMCs).

Main Results:

• Positive isolation achieved up to 89% clearance and 7.6 depletion factor for monocytes labeled with anti-CD14 mAb.
• Monocyte elution yielded 77% of applied cells with 90% purity and >65% viability.
• Negative isolation resulted in up to 81% lymphocyte depletion and 3.8 monocyte enrichment factor with 71% purity and >95% viability.

Conclusions:

• The developed expanded-bed adsorption technique effectively isolates monocytes from human peripheral blood.
• This method demonstrates significant potential for intensifying cell separation processes with high cell purity and viability.
• The combination of expanded-bed adsorption and fluidizable affinity supports offers a promising approach for cell isolation.