Five autoantibodies identified from immune complexes as breast cancer biomarkers

ObjectiveComprehensive identification and profiling of antigens in serum immune complexes (ICs) is crucial for developing early diagnostic biomarkers for cancer. We therefore undertook this study to identify novel IC-derived autoantigens and autoantibodies in patients with breast cancer, and to eval...

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Main Authors: Ningwei Zheng, Yueqi Li, Zhengke Peng, Yaolin Tang, Zhiqiang Liang, Hong Wang, Hong Dai, Gongjun Tan
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1640054/full
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author Ningwei Zheng
Yueqi Li
Zhengke Peng
Yaolin Tang
Zhiqiang Liang
Hong Wang
Hong Dai
Gongjun Tan
author_facet Ningwei Zheng
Yueqi Li
Zhengke Peng
Yaolin Tang
Zhiqiang Liang
Hong Wang
Hong Dai
Gongjun Tan
author_sort Ningwei Zheng
collection DOAJ
description ObjectiveComprehensive identification and profiling of antigens in serum immune complexes (ICs) is crucial for developing early diagnostic biomarkers for cancer. We therefore undertook this study to identify novel IC-derived autoantigens and autoantibodies in patients with breast cancer, and to evaluate their potential as new biomarkers.MethodsICs were purified from serum with C1q and Protein A/G affinity capture. The isolated complexes were digested with papain and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Twelve candidate autoantibodies revealed by LC-MS/MS were first verified with a digital liquid chip method (DLCM) in baseline serum from 40 breast cancer patients and eight healthy controls. Five autoantibodies were then validated in independent cohorts of 33 breast cancer patients and 45 healthy controls, using DLCM.ResultsAutoantibodies targeting PF4, PSMB3, PRPF19, RTCB, SDHA, ENO1, PTBP2, PRDX6, ANP32A, VDAC1, MMP14 and HSPA4 were identified both purification methods. In the verification cohort, IgG autoantibodies against HSPA4, ENO1, PRDX6, PRPF19 and MMP14 were significantly increased in breast cancer patients with areas under the curve (AUCs) of 0.90, 0.89, 0.82, 0.78 and 0.77, respectively. Their combined panel discriminated breast cancer from controls with an AUC of 0.97. In the validation cohort, the same autoantibodies achieved AUCs of 0.79, 0.81, 0.73, 0.87, and 0.82, and the combination of these five autoantibodies yielded an AUC of 0.88.ConclusionsThe autoantibodies identified from ICs can serve as effective serum biomarkers for breast cancer. Anti-HSPA4, anti-PRPF19, anti-ENO1, anti-PRDX6, and anti-MMP14 autoantibodies showed significant increases in breast cancer patients.
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spelling doaj-art-ebe3ca34485147868bc103e2f22e706b2025-08-20T03:31:27ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-07-011610.3389/fimmu.2025.16400541640054Five autoantibodies identified from immune complexes as breast cancer biomarkersNingwei Zheng0Yueqi Li1Zhengke Peng2Yaolin Tang3Zhiqiang Liang4Hong Wang5Hong Dai6Gongjun Tan7Department of Clinical Laboratory, Zhuhai Center for Maternal and Child Healthcare (Zhuhai Women and Children’s Hospital), Zhuhai, ChinaDepartment of Clinical Laboratory, Zhuhai Center for Maternal and Child Healthcare (Zhuhai Women and Children’s Hospital), Zhuhai, ChinaDepartment of Clinical Laboratory, Zhuhai Hospital, Jinan University, Zhuhai, ChinaDepartment of Clinical Laboratory, Zhuhai Center for Maternal and Child Healthcare (Zhuhai Women and Children’s Hospital), Zhuhai, ChinaDepartment of Clinical Laboratory, Zhuhai Center for Maternal and Child Healthcare (Zhuhai Women and Children’s Hospital), Zhuhai, ChinaDepartment of Breast Surgery, Zhuhai Center for Maternal and Child Healthcare (Zhuhai Women and Children’s Hospital), Zhuhai, ChinaDepartment of Gynecology, Zhuhai Center for Maternal and Child Healthcare (Zhuhai Women and Children’s Hospital), Zhuhai, ChinaDepartment of Clinical Laboratory, Zhuhai Center for Maternal and Child Healthcare (Zhuhai Women and Children’s Hospital), Zhuhai, ChinaObjectiveComprehensive identification and profiling of antigens in serum immune complexes (ICs) is crucial for developing early diagnostic biomarkers for cancer. We therefore undertook this study to identify novel IC-derived autoantigens and autoantibodies in patients with breast cancer, and to evaluate their potential as new biomarkers.MethodsICs were purified from serum with C1q and Protein A/G affinity capture. The isolated complexes were digested with papain and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Twelve candidate autoantibodies revealed by LC-MS/MS were first verified with a digital liquid chip method (DLCM) in baseline serum from 40 breast cancer patients and eight healthy controls. Five autoantibodies were then validated in independent cohorts of 33 breast cancer patients and 45 healthy controls, using DLCM.ResultsAutoantibodies targeting PF4, PSMB3, PRPF19, RTCB, SDHA, ENO1, PTBP2, PRDX6, ANP32A, VDAC1, MMP14 and HSPA4 were identified both purification methods. In the verification cohort, IgG autoantibodies against HSPA4, ENO1, PRDX6, PRPF19 and MMP14 were significantly increased in breast cancer patients with areas under the curve (AUCs) of 0.90, 0.89, 0.82, 0.78 and 0.77, respectively. Their combined panel discriminated breast cancer from controls with an AUC of 0.97. In the validation cohort, the same autoantibodies achieved AUCs of 0.79, 0.81, 0.73, 0.87, and 0.82, and the combination of these five autoantibodies yielded an AUC of 0.88.ConclusionsThe autoantibodies identified from ICs can serve as effective serum biomarkers for breast cancer. Anti-HSPA4, anti-PRPF19, anti-ENO1, anti-PRDX6, and anti-MMP14 autoantibodies showed significant increases in breast cancer patients.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1640054/fullcomplement 1Qimmune complex analysisautoantibodiesbreast cancerdigital liquid chip method
spellingShingle Ningwei Zheng
Yueqi Li
Zhengke Peng
Yaolin Tang
Zhiqiang Liang
Hong Wang
Hong Dai
Gongjun Tan
Five autoantibodies identified from immune complexes as breast cancer biomarkers
Frontiers in Immunology
complement 1Q
immune complex analysis
autoantibodies
breast cancer
digital liquid chip method
title Five autoantibodies identified from immune complexes as breast cancer biomarkers
title_full Five autoantibodies identified from immune complexes as breast cancer biomarkers
title_fullStr Five autoantibodies identified from immune complexes as breast cancer biomarkers
title_full_unstemmed Five autoantibodies identified from immune complexes as breast cancer biomarkers
title_short Five autoantibodies identified from immune complexes as breast cancer biomarkers
title_sort five autoantibodies identified from immune complexes as breast cancer biomarkers
topic complement 1Q
immune complex analysis
autoantibodies
breast cancer
digital liquid chip method
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1640054/full
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