Pathogen-activated antibody-secreting cells (ASCs) produce and secrete antigen-specific antibodies. ASCs are detectable in the peripheral blood as early as 3 days after antigen exposure, which makes ASCs a potential biomarker for early disease detection. Here, we present a magnetic capture and detection (MCD) assay for sensitive, on-site detection of ASCs. In this approach, ASCs are enriched through magnetic capture, and secreted antibodies are magnetically detected by a miniaturized nuclear magnetic resonance (μNMR) system. This approach is based entirely on magnetics, which supports high contrast against biological background and simplifies assay procedures. We advanced the MCD system by (i) synthesizing magnetic nanoparticles with high magnetic moments for both cell capture and antibody detection, (ii) developing a miniaturized magnetic device for high-yield cell capture, and (iii) optimizing the μNMR assay for antibody detection. Antibody responses targeting hemolysin E (HlyE) can accurately identify individuals with acute enteric fever. As a proof-of-concept, we applied MCD to detect antibodies produced by HlyE-specific hybridoma cells. The MCD achieved high sensitivity in detecting antibodies secreted from as few as 5 hybridoma cells (50 cells/mL). Importantly, the assay could be performed with whole blood with minimal sample processing.
Bibliographical noteFunding Information:
The authors were supported in part by NIH Grants R33CA202064 (R.W., H.L.), R01HL113156 (H.L.), R21CA205322 (H.L.), R01EB004626 (R.W.), R01EB010011 (R.W.), R33AI100023 (E.T.R., F.Q.), D43 TW005572 (F.K., R.R., T.R.B.), 43TW010362 (T.R.B.); the Massachusetts General Hospital Research Scholar Fund (H.L.); the Massachusetts General Hospital Department of Medicine Transformative Scholars Award (R.C.C.); the Robert Wood Johnson Foundation Harold Amos Medical Faculty Development Program (R.C.C.); the Harvard Medical School DCIP Faculty Fellowship (R.C.C.); the National Research Foundation (NRF) awards by the Ministry of Science, ICT & Future Planning (MSIP) of Korea, 2014R1A6A3A0305972 (K.S.P.) and 2017R1C1B5017724 (K.S.P.); the Massachusetts General Hospital Tosteson Award (K.S.P.); the Institute for Basic Science, IBS-R026-D1 (J.C., H.L.); the IBS Global Postdoctoral Fellowship Award (IBS GPF) (K.S.P.); and the Korea Healthcare Technology R&D Project, Ministry for Health & Welfare Affairs, HI08C2149 (J.C.). The authors thank Prof. Dongwon Yoo for coordinating the IBS GPF, and Dr. Tae-Hyun Shin for helpful comments and discussions.
© 2017 American Chemical Society.
All Science Journal Classification (ASJC) codes
- Materials Science(all)
- Physics and Astronomy(all)