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

Journal: Sensors and Actuators B: Chemical

Article Title: Sequential separation and profiling of extracellular vesicles using antibody-aptamer conjugates

doi: 10.1016/j.snb.2024.136939

Figure Lengend Snippet: Fig. 3. Relative Fluorescence Intensity measured on the spots after the interaction of fluorescently labeled secondary antibody with different AACs (Rabbit IgG-Tag1, Rabbit IgG-Tag2 and Rabbit IgG-Tag3) immobilized on a microarray functionalized with streptavidin (printed at a concentration of 2 mg/mL) and released after incubation with biotin. Blue bars reflect the AAC immobilized on streptavidin, yellow bars represent the AAC left on the spots after biotin-mediated release. Mean ± standard deviation of three independent measurements.

Article Snippet: Supports were spotted using a noncontact microarray spotter (sciFLEXARRAYER S12, Scienion, Berlin) equipped with an 80 μm nozzle.

Techniques: Fluorescence, Labeling, Microarray, Concentration Assay, Incubation, Standard Deviation

Journal: Sensors and Actuators B: Chemical

Article Title: Sequential separation and profiling of extracellular vesicles using antibody-aptamer conjugates

doi: 10.1016/j.snb.2024.136939

Figure Lengend Snippet: Fig. 4. Sequence-selective recapture on DNA microarray of 3 different AACs previously immobilized on streptavidin coated magnetic beads and released upon incubation with biotin. Bars represent the Relative Fluorescence Intensity measured on the spots after the interaction of fluorescently labeled secondary antibody with different AACs. Mean ± standard deviation of three independent measurements.

Article Snippet: Supports were spotted using a noncontact microarray spotter (sciFLEXARRAYER S12, Scienion, Berlin) equipped with an 80 μm nozzle.

Techniques: Sequencing, Microarray, Magnetic Beads, Incubation, Fluorescence, Labeling, Standard Deviation

Journal: Sensors and Actuators B: Chemical

Article Title: Sequential separation and profiling of extracellular vesicles using antibody-aptamer conjugates

doi: 10.1016/j.snb.2024.136939

Figure Lengend Snippet: Fig. 5. Detection of EVs by label free interferometry separated and recaptured on DNA microarray using antiCD9-Tag2 (red bars). In the negative control, AAC- functionalized magnetic beads were incubated only with PBS-M (blue bars). Mean ± standard deviation of three independent measurements.

Article Snippet: Supports were spotted using a noncontact microarray spotter (sciFLEXARRAYER S12, Scienion, Berlin) equipped with an 80 μm nozzle.

Techniques: Microarray, Negative Control, Magnetic Beads, Incubation, Standard Deviation

Journal: Sensors and Actuators B: Chemical

Article Title: Sequential separation and profiling of extracellular vesicles using antibody-aptamer conjugates

doi: 10.1016/j.snb.2024.136939

Figure Lengend Snippet: Fig. 6. Schematic representation of recapture experiment on ExoView R100 (upper panel) and relative high resolution close ups of a Probe2 spot throughout experimental stages (bottom panel). From left to the right it can be seen i) the surface of the DNA spot, ii) label free interferometry picture of EVs captured on the same spot, and iii) red and green fluorescence picture of the same spot after incubation with labeled antibodies. The bar graphs represent label free interferometric and fluorescent detection of EVs on DNA microarray after separation using streptavidin coated magnetic beads: (b) functionalized with antiCD9-Tag2; (c) func- tionalized with Rabbit IgG-Tag2; (d) non functionalized with AACs. As additional negative control, detection of plasma added with antiCD9-Tag2 without previous separation of EVs on magnetic beads (e). Mean ± standard deviation of three independent measurements.

Article Snippet: Supports were spotted using a noncontact microarray spotter (sciFLEXARRAYER S12, Scienion, Berlin) equipped with an 80 μm nozzle.

Techniques: Fluorescence, Incubation, Labeling, Microarray, Magnetic Beads, Negative Control, Clinical Proteomics, Standard Deviation

Journal: Sensors and Actuators B: Chemical

Article Title: Sequential separation and profiling of extracellular vesicles using antibody-aptamer conjugates

doi: 10.1016/j.snb.2024.136939

Figure Lengend Snippet: Fig. 8. Recapture and multiplexed detection on DNA microarray of EVs sepa- rated using antiCD9-Tag2 immobilized on magnetic beads and labeled using antiCD63-Tag1. Mean ± standard deviation of three independent measurements.

Article Snippet: Supports were spotted using a noncontact microarray spotter (sciFLEXARRAYER S12, Scienion, Berlin) equipped with an 80 μm nozzle.

Techniques: Microarray, Magnetic Beads, Labeling, Standard Deviation