TY - JOUR

T1 - An Antibody-Aptamer-Hybrid Lateral Flow Assay for Detection of CXCL9 in Antibody-Mediated Rejection after Kidney Transplantation

AU - Seiler, Lisa K.

AU - Phung, Ngoc Linh

AU - Nikolin, Christoph

AU - Immenschuh, Stephan

AU - Erck, Christian

AU - Kaufeld, Jessica

AU - Haller, Hermann

AU - Falk, Christine S.

AU - Jonczyk, Rebecca

AU - Lindner, Patrick

AU - Thoms, Stefanie

AU - Siegl, Julia

AU - Mayer, Günter

AU - Feederle, Regina

AU - Blume, Cornelia A.

N1 - Funding Information: This work was funded by the start-up project SU02 and the Core Facility Diagnostic Centre of the IFB-Tx (ref. nr. 01EO0802) of Cornelia Blume and the DFG SFB738-B8 projects and the DZIF TTU-IICH07.801 of Christine Falk. The continued biomarker evaluation and new establishment of the LFA was funded by the European Regional Development Fund (ERDF), subproject A (ref. nr. 85006385) and Central Innovation Program for Small and Medium Entrepreneurs of the Federal Ministry for Economic Affairs and Energy Germany, (ref. nr. ZF4409702 CR8). We are grateful for funding by the Deutsche Forschungsgemeinschaft (Center of Aptamer Research and Development (3442/7-1)) to Günter Mayer. Funding Information: Funding: This work was funded by the start-up project SU02 and the Core Facility Diagnostic Centre of the IFB-Tx (ref. nr. 01EO0802) of Cornelia Blume and the DFG SFB738-B8 projects and the DZIF TTU-IICH07.801 of Christine Falk. The continued biomarker evaluation and new establishment of the LFA was funded by the European Regional Development Fund (ERDF), subproject A (ref. nr. 85006385) and Central Innovation Program for Small and Medium Entrepreneurs of the Federal Ministry for Economic Affairs and Energy Germany, (ref. nr. ZF4409702 CR8). We are grateful for funding by the Deutsche Forschungsgemeinschaft (Center of Aptamer Research and Development (3442/7-1)) to Günter Mayer.

PY - 2022/1/25

Y1 - 2022/1/25

N2 - Chronic antibody-mediated rejection (AMR) is a key limiting factor for the clinical outcome of a kidney transplantation (Ktx), where early diagnosis and therapeutic intervention is needed. This study describes the identification of the biomarker CXC-motif chemokine ligand (CXCL) 9 as an indicator for AMR and presents a new aptamer-antibody-hybrid lateral flow assay (hybrid-LFA) for detection in urine. Biomarker evaluation included two independent cohorts of kidney transplant recipients (KTRs) from a protocol biopsy program and used subgroup comparisons according to BANFF-classifications. Plasma, urine and biopsy lysate samples were analyzed with a Luminex-based multiplex assay. The CXCL9-specific hybrid-LFA was developed based upon a specific rat antibody immobilized on a nitrocellulose-membrane and the coupling of a CXCL9-binding aptamer to gold nanoparticles. LFA performance was assessed according to receiver operating characteristic (ROC) analysis. Among 15 high-scored biomarkers according to a neural network analysis, significantly higher levels of CXCL9 were found in plasma and urine and biopsy lysates of KTRs with biopsyproven AMR. The newly developed hybrid-LFA reached a sensitivity and specificity of 71% and an AUC of 0.79 for CXCL9. This point-of-care-test (POCT) improves early diagnosis-making in AMR after Ktx, especially in KTRs with undetermined status of donor-specific HLA-antibodies.

AB - Chronic antibody-mediated rejection (AMR) is a key limiting factor for the clinical outcome of a kidney transplantation (Ktx), where early diagnosis and therapeutic intervention is needed. This study describes the identification of the biomarker CXC-motif chemokine ligand (CXCL) 9 as an indicator for AMR and presents a new aptamer-antibody-hybrid lateral flow assay (hybrid-LFA) for detection in urine. Biomarker evaluation included two independent cohorts of kidney transplant recipients (KTRs) from a protocol biopsy program and used subgroup comparisons according to BANFF-classifications. Plasma, urine and biopsy lysate samples were analyzed with a Luminex-based multiplex assay. The CXCL9-specific hybrid-LFA was developed based upon a specific rat antibody immobilized on a nitrocellulose-membrane and the coupling of a CXCL9-binding aptamer to gold nanoparticles. LFA performance was assessed according to receiver operating characteristic (ROC) analysis. Among 15 high-scored biomarkers according to a neural network analysis, significantly higher levels of CXCL9 were found in plasma and urine and biopsy lysates of KTRs with biopsyproven AMR. The newly developed hybrid-LFA reached a sensitivity and specificity of 71% and an AUC of 0.79 for CXCL9. This point-of-care-test (POCT) improves early diagnosis-making in AMR after Ktx, especially in KTRs with undetermined status of donor-specific HLA-antibodies.

KW - Antibody

KW - Antibody-mediated rejection (AMR)

KW - Aptamer

KW - Aptamer-antibody-hybrid lateral flow assay (hybrid-LFA)

KW - Biomarkers

KW - CXCL9 (MIG)

KW - Neural net analysis

UR - http://www.scopus.com/inward/record.url?scp=85124327642&partnerID=8YFLogxK

U2 - 10.3390/diagnostics12020308

DO - 10.3390/diagnostics12020308

M3 - Article

AN - SCOPUS:85124327642

VL - 12

JO - Diagnostics

JF - Diagnostics

SN - 2075-4418

IS - 2

M1 - 308

ER -