Rapid capture and labeling of cells on single domain antibodies-functionalized flow cell.
Current techniques to characterize leukocyte subgroups in blood require long sample preparation times and sizable sample volumes. A simplified method for leukocyte characterization using smaller blood volumes would thus be useful in diagnostic settings. Here we describe a flow system comprised of two functionalized graphene oxide (GO) surfaces that allow the capture of distinct leukocyte populations from small volumes blood using camelid single-domain antibodyfragments (VHHs) as capture agents. We used site-specifically labeled leukocytes to detect and identify cells exposed to fungal challenge. Combining the chemical and optical properties of GO with the versatility of the VHH scaffold in the context of a flow system provides a quick and efficient method for the capture and characterization of functional leukocytes.
Whitehead Institute for Biomedical Research, Cambridge, MA, USA; Institute of Biomedical Engineering, National Chiao Tung University, Hsinchu 30010, Taiwan; Department of Biological Science and Technology, National Chiao Tung University, Hsinchu 30010, Taiwan.
Guan-Yu
GY
Chen
Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
Zeyang
Z
Li
Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
Joao N
JN
Duarte
Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
Alexandre
A
Esteban
Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
Ross W
RW
Cheloha
Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
Christopher S
CS
Theile
Whitehead Institute for Biomedical Research, Cambridge, MA, USA; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.
Gerald R
GR
Fink
Whitehead Institute for Biomedical Research, Cambridge, MA, USA; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. Electronic address: ploegh@wi.mit.edu.
Hidde L
HL
Ploegh
D019881:Aminoacyltransferases / Q000032:analysis; D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry*; D001426:Bacterial Proteins / Q000032:analysis; D002176:Candida albicans / Q000302:isolation & purification*; D002177:Candidiasis / Q000097:blood; D002469:Cell Separation / Q000379:methods*; D003546:Cysteine Endopeptidases / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D006108:Graphite / Q000737:chemistry*; D007962:Leukocytes / Q000166:cytology* / Q000382:microbiology*; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D049329:Nanostructures / Q000737:chemistry; D061905:Single-Domain Antibodies / Q000737:chemistry*
D055910:Antibodies, Immobilized; D001426:Bacterial Proteins; D061905:Single-Domain Antibodies; D006108:Graphite; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases
Cell detection; Enzymatic labeling; Graphene oxide; Leukocytes; Single domain antibody; Sortase