Centrifugo-Magnetophoretic Purification of CD4+ Cells from Whole Blood Toward Future HIV/AIDS Point-of-Care Applications

J Lab Autom. 2014 Jun;19(3):285-96. doi: 10.1177/2211068213504759. Epub 2013 Sep 20.

Abstract

In medical diagnostics, detection of cells exhibiting specific phenotypes constitutes a paramount challenge. Detection technology must ensure efficient isolation of (often rare) targets while eliminating nontarget background cells. Technologies exist for such investigations, but many require high levels of expertise, expense, and multistep protocols. Increasing automation, miniaturization, and availability of such technologies is an aim of microfluidic lab-on-a-chip strategies. To this end, we present an integrated, dual-force cellular separation strategy using centrifugo-magnetophoresis. Whole blood spiked with target cells is incubated with (super-)paramagnetic microparticles that specifically bind phenotypic markers on target cells. Under rotation, all cells sediment into a chamber located opposite a co-rotating magnet. Unbound cells follow the radial vector, but under the additional attraction of the lateral magnetic field, bead-bound target cells are deflected to a designated reservoir. This multiforce separation is continuous and low loss. We demonstrate separation efficiently up to 92% for cells expressing the HIV/AIDS relevant epitope (CD4) from whole blood. Such highly selective separation systems may be deployed for accurate diagnostic cell isolations from biological samples such as blood. Furthermore, this high efficiency is delivered in a cheap and simple device, thus making it an attractive option for future deployment in resource-limited settings.

Keywords: HIV/AIDS diagnostics; cell purification; cell separation; centrifugal microfluidics; lab-on-a-disc.

Publication types

  • Comparative Study
  • Validation Study

MeSH terms

  • Acquired Immunodeficiency Syndrome / diagnosis
  • Acquired Immunodeficiency Syndrome / economics
  • Acquired Immunodeficiency Syndrome / immunology
  • Acquired Immunodeficiency Syndrome / pathology
  • Automation, Laboratory / economics
  • Automation, Laboratory / instrumentation*
  • Blood Buffy Coat / cytology
  • Blood Buffy Coat / immunology
  • Blood Buffy Coat / metabolism
  • Blood Buffy Coat / pathology
  • CD4-Positive T-Lymphocytes / cytology*
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / metabolism
  • CD4-Positive T-Lymphocytes / pathology
  • Centrifugation / economics
  • Centrifugation / instrumentation
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / chemistry
  • HIV Infections / diagnosis
  • HIV Infections / economics
  • HIV Infections / immunology
  • HIV Infections / pathology
  • HL-60 Cells
  • HeLa Cells
  • Health Care Costs
  • Humans
  • Immunomagnetic Separation / economics
  • Immunomagnetic Separation / instrumentation*
  • Lab-On-A-Chip Devices* / economics
  • Microfluidics / economics
  • Microfluidics / instrumentation*
  • Microspheres
  • Point-of-Care Systems* / economics
  • Point-of-Care Testing / economics
  • Proof of Concept Study
  • Spectrometry, Fluorescence / economics
  • Spectrometry, Fluorescence / instrumentation

Substances

  • Fluorescent Dyes