| Size | 1 slide (16 wells), 2 slides (32 wells) |
| Estimated Lead Time | 1-2 business days |
| Species | Human |
| Compatible Sample Types | Plasma, Serum, Ascites, Hybridoma cell culture medium, Purified Antibodies, Purified Proteins |
| Protein Name / Synonyms | A29L, A30L, A33R, A35R, B16R, C19L (identical to vaccinia virus F13L), D6L, E8L, H3L, I1L, L1R |
| Quantitative/Semi-Quantitative | Semi-Quantitative |
| Number of Targets Detected | 11 |
| Solid Support | Glass Slide |
| Method Of Detection | Fluorescence Laser Scanner |
| Research Area | Infectious Disease, Mpox |
| Shipping Type | Blue ice |
| Storage | -20°C |
Recombinant Monkeypox virus (MPXV) proteins derived from bacterial and human cells (HEK293) are spotted in triplicate on a standard glass slide (25 mm x 75 mm x 1 mm), along with positive and negative controls to monitor each incubation step. When liquid sample types, such as serum or purified antibodies, are incubated on the array, "primary" antibodies in the sample will bind to their specific target proteins. Specific primary antibody isotypes (IgG, IgM, or IgA) from serum, plasma, purified antibodies, etc. are then targeted by biotin-conjugated anti-IgG, IgM, or IgA secondary antibodies, respectively, that are specific for the host species.
A fluorophore-conjugated streptavidin molecule is then added, which binds to the biotin on the secondary antibody. The fluorophore enables the detection of the immobilized primary antibody via fluorescence using a laser scanner. The fluorescence signal is proportional to the amount of immobilized antibody. Since each spot represents a unique protein that is known, the specific target(s) bound by the antibodies can be ascertained.
Three serum samples (Serum #1, #2, and #3) from Monkeypox virus infected individuals (June 2022, USA) were assayed using the Raybio® Monkeypox Virus Protein Array G1 (Cat # PAV-MPHG-G1). Three isoforms — IgG, IgM and IgA were simultaneously probed from each sample, revealing a significant immune response to various viral target proteins (Figure 1, Table 1). Interestingly, both bacterial-derived and human cell-derived recombinant viral proteins had similar antibody responses.
Figure 1. Monkeypox Virus Protein Array G1 run with patient serum samples. Monkeypox Virus Protein Array G1 was printed on a glass-slide array with Bio-BSA and IgG/IgM/IgA positive controls (white and red highlighted boxes, respectively) and 18 E. coli- or HEK293-derived Monkeypox viral proteins (Table 1). Serum samples from three Monkeypox virus infected individuals (Serum #1, #2, and #3) were obtained, diluted 1,000-fold and incubated in the array along with a blocking buffer negative control. The arrays were probed using biotin-conjugated anti-human IgG, IgM, and IgA, respectively. Followed by wash steps, the array was incubated with Cy3 equivalent dye-conjugated streptavidin. The array signals were visualized using a laser scanner capable of measuring signals in the green channel.
Table 1. Immune response of Monkeypox virus infected patients to different MPXV proteins
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Monkeypox Virus Protein Array G1
Monkeypox Virus Protein Array G1