protoarray® protein microarray (v.1.0)  (Promega)

Journal: Journal of Virology

Article Title: An African swine fever virus-specific antibody reactome reveals antigens as potential candidates for vaccine development

doi: 10.1128/jvi.00478-25

Figure Lengend Snippet: Construction and characterization of the ASFV proteome microarray. In parallel with the controls, a total of 160 GST-tagged ASFV proteins were printed in triplicate onto a PATH substrate slide, generating 14 identical subarrays per slide. The slides were probed with an anti-GST antibody and a Cy3-labeled secondary antibody. (a) A representative subarray (upper) and the layout of the array (lower). (b) Histogram analysis of the fluorescence intensity of all the immobilized ASFV proteins based on their N-terminal GST tag probed with an anti-GST antibody and a fluorescently labeled secondary antibody. (c) Fluorescence intensity distribution of the spots of ASFV proteins and negative controls (including blank, BSA, IgG, IgM, and IgA) after probing with an anti-GST antibody and a fluorescently labeled secondary antibody. (d) Representative subarrays probed with sera from an ASFV-infected pig and a healthy pig. The IgG signals are shown in green. (e) Correlation analysis showing the repeated experimental results for the same serum sample.

Article Snippet: Briefly, using a Super Marathon printer (Arrayjet, UK), identical protein arrays in a 2 × 7 subarray format were generated by printing affinity-purified 160 ASFV proteins, accompanied by negative (BSA and GST) and positive controls (anti-swine IgG [Novus Biologicals, USA, Cat# NBP1-97054], IgM [Novus Biologicals, USA, Cat# NBP1-96788], and IgA [Alpha Diagnostic International, USA, Cat# 20017-4-1]) and land markers, in triplicate, on PATH Protein Microarray Slides (GraceBio-Labs, Oregon, USA).

Techniques: Microarray, Labeling, Fluorescence, Infection

Journal: Journal of Virology

Article Title: An African swine fever virus-specific antibody reactome reveals antigens as potential candidates for vaccine development

doi: 10.1128/jvi.00478-25

Figure Lengend Snippet: High-throughput analysis of sera from ASFV-infected pigs using a proteome microarray. (a) Timelines of animal treatment and sample collection. Group A pigs ( n = 5) were infected with a virulent ASFV CN/GS 2018 strain at 1 HAD 50 . Three pigs died by 15 dpi, while the other two pigs developed clinical symptoms of ASF during the observation period but ultimately survived. Serum samples were collected at 3, 5, 7, 9, and 15 dpi. Group B pigs ( n = 5) inoculated with 10 4 HAD 50 of ASFV-GS-ΔMGF360-18R/DP71L/DP96R survived the 17-day observation period and remained alive after being challenged with 10 2 HAD 50 of the parental virus during the 13-day observation period. Serum samples were collected at 0, 7, 13, and 17 dpi and at 7 dpc. Group C pigs inoculated with 10 4 HAD 50 of ASFV-GS-ΔMGF110/360-9L ( n = 6) survived the 17-day observation period and remained alive after being challenged with 10 2 HAD 50 of the parental virus during the 13-day observation period. Serum samples were collected at 7, 13, and 17 dpi and 7 dpc. (b) Uniform manifold approximation and projection (UMAP) of the 160-ASFV-protein-specific IgG signals in the serum samples from the three groups. Each point represents an individual serum sample. (c) The amounts of the IgG-positive ASFV proteins and the shared portion from groups B and C.

Article Snippet: Briefly, using a Super Marathon printer (Arrayjet, UK), identical protein arrays in a 2 × 7 subarray format were generated by printing affinity-purified 160 ASFV proteins, accompanied by negative (BSA and GST) and positive controls (anti-swine IgG [Novus Biologicals, USA, Cat# NBP1-97054], IgM [Novus Biologicals, USA, Cat# NBP1-96788], and IgA [Alpha Diagnostic International, USA, Cat# 20017-4-1]) and land markers, in triplicate, on PATH Protein Microarray Slides (GraceBio-Labs, Oregon, USA).

Techniques: High Throughput Screening Assay, Infection, Microarray, Virus