Journal: Frontiers in Immunology

Article Title: Stage-associated immunoproteomic profiling of serum autoantibody-captured retinal antigens in age-related macular degeneration

doi: 10.3389/fimmu.2026.1824524

Figure Lengend Snippet: Overview of the experimental workflow for analyzing retinal antigen profiles captured by serum autoantibodies from patients with different stages of AMD. Antibodies isolated from AMD patient serum were captured on Protein G magnetic beads and incubated overnight with porcine retinal lysates. Unbound retinal antigens were washed away, and antibody-bound antigens were subsequently subjected to MS analysis. Selected differentially expressed retinal antigen targets identified by MS were further evaluated and validated in serum and tear samples from AMD patients using antigen microarray technology. The image was created using BioRender.com ( https://biorender.com/ , accessed on 14. June 2026).

Article Snippet: Commercially obtained antigens ( ) were spotted in triplicates onto nitrocellulose-coated slides (Oncyte, Grace Bio Labs, USA) using a non-contact microarray spotter (sciFLEXARRAYER S3, Scienion, Germany).

Techniques: Isolation, Magnetic Beads, Incubation, Microarray

Journal: Frontiers in Immunology

Article Title: Stage-associated immunoproteomic profiling of serum autoantibody-captured retinal antigens in age-related macular degeneration

doi: 10.3389/fimmu.2026.1824524

Figure Lengend Snippet: Selection and validation of candidate retinal antigens recognized by serum and tear autoantibodies in patients with different stages of AMD. (A) Venn diagram showing shared and stage-specific differentially enriched retinal antigens captured by serum autoantibodies across AMD subgroups. (B) Heatmap showing the relative abundance of selected candidate retinal antigens across AMD stages. Red indicates increased abundance and blue indicates decreased abundance. Candidate antigens selected for further validation are highlighted with red boxes. (C–H) Antigen microarray validation of serum and tear autoantibody reactivity against selected retinal antigens, including PSMD6, HSPA12A, PTGES3, ATP5A1, PFKM, and SDHB, in CTRL and AMD subgroups. Data are presented as mean ± SE. *p < 0.05.

Article Snippet: Commercially obtained antigens ( ) were spotted in triplicates onto nitrocellulose-coated slides (Oncyte, Grace Bio Labs, USA) using a non-contact microarray spotter (sciFLEXARRAYER S3, Scienion, Germany).

Techniques: Selection, Biomarker Discovery, Microarray

Journal: Frontiers in Immunology

Article Title: Stage-associated immunoproteomic profiling of serum autoantibody-captured retinal antigens in age-related macular degeneration

doi: 10.3389/fimmu.2026.1824524

Figure Lengend Snippet: Validation and overlap analysis of selected candidate retinal antigens recognized by serum and tear autoantibodies in patients with different stages of AMD. (A) Venn diagram showing shared and stage-specific differentially enriched retinal antigens captured by serum autoantibodies across AMD subgroups. (B–G) A subset of candidate retinal antigen targets identified by serum autoantibody-based proteomic profiling, including FTH1, SAG, VIM, PRMT5, SEC22B, and HIST2H2AA3, was selected for further validation by antigen microarray analysis in serum and tear samples from CTRL and AMD subgroups. Data are presented as mean ± SE, with whiskers indicating 1.96 × SE.*p < 0.05. (H) Venn diagram showing the overlap among serum proteomics-positive results, serum autoantibody microarray-positive results, and tear autoantibody microarray-positive results. The antigen microarray analysis was performed as a targeted validation of selected MS-derived retinal antigen targets. Candidate antigens supported by multiple datasets are highlighted in red.

Article Snippet: Commercially obtained antigens ( ) were spotted in triplicates onto nitrocellulose-coated slides (Oncyte, Grace Bio Labs, USA) using a non-contact microarray spotter (sciFLEXARRAYER S3, Scienion, Germany).

Techniques: Biomarker Discovery, Microarray, Derivative Assay