Clinical microfluidics for neutrophil genomics and proteomics

• Published in: Nature medicine Vol. 16; no. 9; pp. 1042 - 1047
• Main Authors: Kotz, Kenneth T, Xiao, Wenzong, Miller-Graziano, Carol, Qian, Wei-Jun, Russom, Aman, Warner, Elizabeth A, Moldawer, Lyle L, De, Asit, Bankey, Paul E, Petritis, Brianne O, Camp, David G, Rosenbach, Alan E, Goverman, Jeremy, Fagan, Shawn P, Brownstein, Bernard H, Irimia, Daniel, Xu, Weihong, Wilhelmy, Julie, Mindrinos, Michael N, Smith, Richard D, Davis, Ronald W, Tompkins, Ronald G, Toner, Mehmet
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.09.2010; Nature Publishing Group
• Subjects: 631/250/2504/223/1699; 631/337/475; 639/638/11/877; Analysis; Antibodies, Monoclonal; Antigens, CD - genetics; Antigens, CD - immunology; Biomedical and Life Sciences; Biomedicine; Biotinylation; Burns - physiopathology; Cancer Research; Cell Adhesion Molecules - genetics; Cell Adhesion Molecules - immunology; DNA - genetics; DNA - isolation & purification; Gene expression; Genomics; Genomics - methods; GPI-Linked Proteins; Health aspects; Humans; Immune response; Infectious Diseases; Messenger RNA; Metabolic Diseases; Microfluidics - methods; Miniaturization; Molecular Medicine; Neurosciences; Neutrophils; Neutrophils - cytology; Neutrophils - physiology; Oligonucleotide Array Sequence Analysis; Proteomics; Proteomics - methods; RNA - genetics; RNA - isolation & purification; Semiconductors; technical-report; Validation studies; Wounds and Injuries - physiopathology
• ISSN: 1078-8956; 1546-170X; 1546-170X
• DOI: 10.1038/nm.2205

Standard methods of neutrophil isolation require skilled personnel, are time consuming and use large blood volumes. Kotz and his colleagues have developed a rapid microfluidic chip-based approach for rapidly isolating neutrophils directly from whole blood with 'on-chip' processing for mRNA and protein isolation. The device, which yields sufficient quantities and purities for downstream genomic or proteomic analysis, was validated in a multicenter clinical study of the immune response to severe trauma and burn injury. Neutrophils have key roles in modulating the immune response. We present a robust methodology for rapidly isolating neutrophils directly from whole blood with 'on-chip' processing for mRNA and protein isolation for genomics and proteomics. We validate this device with an ex vivo stimulation experiment and by comparison with standard bulk isolation methodologies. Last, we implement this tool as part of a near-patient blood processing system within a multi-center clinical study of the immune response to severe trauma and burn injury. The preliminary results from a small cohort of subjects in our study and healthy controls show a unique time-dependent gene expression pattern clearly demonstrating the ability of this tool to discriminate temporal transcriptional events of neutrophils within a clinical setting.