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(A) Screening burden reduction: For three independent targets <t>(TREM2,</t> CHI3L1, and CD28), HTS-Oracle reduced the number of compounds requiring experimental testing by >99% for TREM2 and CHI3L1, and by 70% for CD28, relative to traditional HTS workflows. (B) Hit rate improvement: HTS-Oracle achieved substantial increase in hit rate when compared with traditional HTS, including 30.6× and 87.5× for TREM2 (single-dose and validated datasets, respectively), 176× for CHI3L1 (single-dose), and 8.4× for CD28.
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(A) Screening burden reduction: For three independent targets <t>(TREM2,</t> CHI3L1, and CD28), HTS-Oracle reduced the number of compounds requiring experimental testing by >99% for TREM2 and CHI3L1, and by 70% for CD28, relative to traditional HTS workflows. (B) Hit rate improvement: HTS-Oracle achieved substantial increase in hit rate when compared with traditional HTS, including 30.6× and 87.5× for TREM2 (single-dose and validated datasets, respectively), 176× for CHI3L1 (single-dose), and 8.4× for CD28.
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(A) Screening burden reduction: For three independent targets (TREM2, CHI3L1, and CD28), HTS-Oracle reduced the number of compounds requiring experimental testing by >99% for TREM2 and CHI3L1, and by 70% for CD28, relative to traditional HTS workflows. (B) Hit rate improvement: HTS-Oracle achieved substantial increase in hit rate when compared with traditional HTS, including 30.6× and 87.5× for TREM2 (single-dose and validated datasets, respectively), 176× for CHI3L1 (single-dose), and 8.4× for CD28.

Journal: bioRxiv

Article Title: HTS-Oracle: Experimentally validated AI-enabled prioritization for generalizable small molecule hit discovery

doi: 10.1101/2025.11.26.690784

Figure Lengend Snippet: (A) Screening burden reduction: For three independent targets (TREM2, CHI3L1, and CD28), HTS-Oracle reduced the number of compounds requiring experimental testing by >99% for TREM2 and CHI3L1, and by 70% for CD28, relative to traditional HTS workflows. (B) Hit rate improvement: HTS-Oracle achieved substantial increase in hit rate when compared with traditional HTS, including 30.6× and 87.5× for TREM2 (single-dose and validated datasets, respectively), 176× for CHI3L1 (single-dose), and 8.4× for CD28.

Article Snippet: His-tagged TREM2 (SinoBiological, #11084-H08H) was labeled with RED-tris-NTA 2 nd generation dye (NanoTemper Technologies, #MO-L018) according to the manufacturer’s instructions and the protein concentration was adjusted to 40 nM.

Techniques:

(A) Results of single-dose screening for TREM2 using the Fnorm readout after 5 seconds. Negative control (DMSO in assay buffer) and positive control ( T2337 ) are shown as green dots, hits outside of a ten standard deviation range (shown as orange area) are displayed as blue dots. Compounds that had to be excluded after control experiments (Figure S3) are indicated as red hollow dots. Results from two independent experiments, graph created with GraphPad Prism 10. (B) Dose-response binding curves for validated TREM2 hits. Compounds were tested across an 8-point concentration series (200 µM to 0.005 µM) using Monolith X to determine binding affinities (n=2). (C) Individual dose-response binding isotherms for the seven validated TREM2 modulators. (D) Overview of HTS-Oracle workflow and performance metrics for TREM2 hit discovery.

Journal: bioRxiv

Article Title: HTS-Oracle: Experimentally validated AI-enabled prioritization for generalizable small molecule hit discovery

doi: 10.1101/2025.11.26.690784

Figure Lengend Snippet: (A) Results of single-dose screening for TREM2 using the Fnorm readout after 5 seconds. Negative control (DMSO in assay buffer) and positive control ( T2337 ) are shown as green dots, hits outside of a ten standard deviation range (shown as orange area) are displayed as blue dots. Compounds that had to be excluded after control experiments (Figure S3) are indicated as red hollow dots. Results from two independent experiments, graph created with GraphPad Prism 10. (B) Dose-response binding curves for validated TREM2 hits. Compounds were tested across an 8-point concentration series (200 µM to 0.005 µM) using Monolith X to determine binding affinities (n=2). (C) Individual dose-response binding isotherms for the seven validated TREM2 modulators. (D) Overview of HTS-Oracle workflow and performance metrics for TREM2 hit discovery.

Article Snippet: His-tagged TREM2 (SinoBiological, #11084-H08H) was labeled with RED-tris-NTA 2 nd generation dye (NanoTemper Technologies, #MO-L018) according to the manufacturer’s instructions and the protein concentration was adjusted to 40 nM.

Techniques: Negative Control, Positive Control, Standard Deviation, Control, Binding Assay, Concentration Assay

Primary screening of compounds at 250 μM (with 3.1% DMSO). Green, brown, black, and orange indicate potential hits, negative control/reference (buffer with 3.1% DMSO), non-binders, and positive control ( G28 ), respectively; (B). (C) Overview of HTS-Oracle workflow and performance metrics for TREM2 hit discovery.

Journal: bioRxiv

Article Title: HTS-Oracle: Experimentally validated AI-enabled prioritization for generalizable small molecule hit discovery

doi: 10.1101/2025.11.26.690784

Figure Lengend Snippet: Primary screening of compounds at 250 μM (with 3.1% DMSO). Green, brown, black, and orange indicate potential hits, negative control/reference (buffer with 3.1% DMSO), non-binders, and positive control ( G28 ), respectively; (B). (C) Overview of HTS-Oracle workflow and performance metrics for TREM2 hit discovery.

Article Snippet: His-tagged TREM2 (SinoBiological, #11084-H08H) was labeled with RED-tris-NTA 2 nd generation dye (NanoTemper Technologies, #MO-L018) according to the manufacturer’s instructions and the protein concentration was adjusted to 40 nM.

Techniques: Negative Control, Positive Control

(A) Screening burden reduction: For three independent targets (TREM2, CHI3L1, and CD28), HTS-Oracle reduced the number of compounds requiring experimental testing by >99% for TREM2 and CHI3L1, and by 70% for CD28, relative to traditional HTS workflows. (B) Hit rate improvement: HTS-Oracle achieved substantial increase in hit rate when compared with traditional HTS, including 30.6× and 87.5× for TREM2 (single-dose and validated datasets, respectively), 176× for CHI3L1 (single-dose), and 8.4× for CD28.

Journal: bioRxiv

Article Title: HTS-Oracle: Experimentally validated AI-enabled prioritization for generalizable small molecule hit discovery

doi: 10.1101/2025.11.26.690784

Figure Lengend Snippet: (A) Screening burden reduction: For three independent targets (TREM2, CHI3L1, and CD28), HTS-Oracle reduced the number of compounds requiring experimental testing by >99% for TREM2 and CHI3L1, and by 70% for CD28, relative to traditional HTS workflows. (B) Hit rate improvement: HTS-Oracle achieved substantial increase in hit rate when compared with traditional HTS, including 30.6× and 87.5× for TREM2 (single-dose and validated datasets, respectively), 176× for CHI3L1 (single-dose), and 8.4× for CD28.

Article Snippet: The single-dose screening was conducted as described previously using recombinant human His-tagged TREM2 (SinoBiological, #11084-H08H) labeled with RED-tris-NTA 2 nd generation dye (NanoTemper Technologies, #MO-L018).

Techniques:

(A) Results of single-dose screening for TREM2 using the Fnorm readout after 5 seconds. Negative control (DMSO in assay buffer) and positive control ( T2337 ) are shown as green dots, hits outside of a ten standard deviation range (shown as orange area) are displayed as blue dots. Compounds that had to be excluded after control experiments (Figure S3) are indicated as red hollow dots. Results from two independent experiments, graph created with GraphPad Prism 10. (B) Dose-response binding curves for validated TREM2 hits. Compounds were tested across an 8-point concentration series (200 µM to 0.005 µM) using Monolith X to determine binding affinities (n=2). (C) Individual dose-response binding isotherms for the seven validated TREM2 modulators. (D) Overview of HTS-Oracle workflow and performance metrics for TREM2 hit discovery.

Journal: bioRxiv

Article Title: HTS-Oracle: Experimentally validated AI-enabled prioritization for generalizable small molecule hit discovery

doi: 10.1101/2025.11.26.690784

Figure Lengend Snippet: (A) Results of single-dose screening for TREM2 using the Fnorm readout after 5 seconds. Negative control (DMSO in assay buffer) and positive control ( T2337 ) are shown as green dots, hits outside of a ten standard deviation range (shown as orange area) are displayed as blue dots. Compounds that had to be excluded after control experiments (Figure S3) are indicated as red hollow dots. Results from two independent experiments, graph created with GraphPad Prism 10. (B) Dose-response binding curves for validated TREM2 hits. Compounds were tested across an 8-point concentration series (200 µM to 0.005 µM) using Monolith X to determine binding affinities (n=2). (C) Individual dose-response binding isotherms for the seven validated TREM2 modulators. (D) Overview of HTS-Oracle workflow and performance metrics for TREM2 hit discovery.

Article Snippet: The single-dose screening was conducted as described previously using recombinant human His-tagged TREM2 (SinoBiological, #11084-H08H) labeled with RED-tris-NTA 2 nd generation dye (NanoTemper Technologies, #MO-L018).

Techniques: Negative Control, Positive Control, Standard Deviation, Control, Binding Assay, Concentration Assay

Primary screening of compounds at 250 μM (with 3.1% DMSO). Green, brown, black, and orange indicate potential hits, negative control/reference (buffer with 3.1% DMSO), non-binders, and positive control ( G28 ), respectively; (B). (C) Overview of HTS-Oracle workflow and performance metrics for TREM2 hit discovery.

Journal: bioRxiv

Article Title: HTS-Oracle: Experimentally validated AI-enabled prioritization for generalizable small molecule hit discovery

doi: 10.1101/2025.11.26.690784

Figure Lengend Snippet: Primary screening of compounds at 250 μM (with 3.1% DMSO). Green, brown, black, and orange indicate potential hits, negative control/reference (buffer with 3.1% DMSO), non-binders, and positive control ( G28 ), respectively; (B). (C) Overview of HTS-Oracle workflow and performance metrics for TREM2 hit discovery.

Article Snippet: The single-dose screening was conducted as described previously using recombinant human His-tagged TREM2 (SinoBiological, #11084-H08H) labeled with RED-tris-NTA 2 nd generation dye (NanoTemper Technologies, #MO-L018).

Techniques: Negative Control, Positive Control