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Biosens. Bioelectron., Feb 06 2025, 275 S0956-5663(25)00109-5 | https://doi.org/10.1016/j.bios.2025.117235 Novel Siglec-1/β-CD/rGO impedimetric biosensor for detecting Neu5Ac expression in red blood cells

Duc-Trung Pham ^a,1, Thi Xoan Hoang ^b,1, Jaehwan Park ^a,c, My-Van Tieu ^a, Tra My Tran ^b, Sungho Park ^a,c, Sathish Panneer Selvam ^a, Hyeon Ho Lee ^b, Ji Eun Park ^e,f, Myung Jin Oh ^e,f, Hyun Joo An ^e,f, Jae Young Kim ^b, Sungbo Cho ^a,c,d

^aDepartment of Electronic Engineering, Gachon University, Seongnam-si, Gyeonggi-do, 13120, Republic of Korea
^bDepartment of Life Science, Gachon University, Seongnam-si, Gyeonggi-do, 13120, Republic of Korea
^cDepartment of Semiconductor Engineering, Gachon University, Seongnam-si, Gyeonggi-do, 13120, Republic of Korea
^dDepartment of Health Sciences and Technology, GAIHST, Gachon University, Incheon, 21999, Republic of Korea
^eGraduate School of Analytical Science and Technology, Chungnam National University, Daejeon, 34134, Republic of Korea
^fAsia Glycomics Reference Site, Daejeon, 34134, Republic of Korea

¹The co-first authors Duc-Trung Pham and Thi Xoan Hoang contributed equally to this work.
Corresponding authors: Jae Young Kim, Sungbo Cho

Abstract

This study introduces a beta-cyclodextrin/reduced graphene oxide/indium tin oxide (β-CD/rGO/ITO) microelectrode platform for the real-time, non-invasive, and label-free detection of N-acetylneuraminic acid (Neu5Ac) on red blood cell (RBC) surfaces. By leveraging advances in CRISPR-Cas9 technology, genetically modified pig-derived RBCs with reduced immunogenicity have been developed for xenotransfusion applications. However, elevated Neu5Ac expression in some modified RBCs complicates immunological assessment. The developed platform demonstrated exceptional sensitivity, specificity, and reusability, enabling selective monitoring of Neu5Ac levels on RBC surfaces through interactions with Siglec-1, a macrophage receptor for Neu5Ac. Results effectively distinguished genetically modified pig RBCs (double knockout (DKO), triple knockout (TKO), and quadruple knockout (QKO)) from human RBCs (HuRBCs) based on Neu5Ac expression. DKO RBCs exhibited the highest impedance value (50.43 ± 3.28 Ω), while HuRBCs showed the lowest (6.29 ± 3.14 Ω). The platform's 98.6% reusability after sialidase treatment and its ability to screen substances that inhibit Neu5Ac-mediated immune recognition highlight its potential for therapeutic applications. These features establish the β-CD/rGO/ITO microelectrode as a valuable tool for quality assessment of xenogeneic RBCs and for advancing clinical and translational applications in xenotransfusion medicine.

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