A35R Antibody Bioactivity Verification

Using ELISA, the Monkeypox virus A35R antigen (C1620, C1621) was used to detect MPXV A35R monoclonal antibodies R383e5, R387c6, R386t7, R384c2, and R392a3. The EC50 values for R383e5 and R387c6 were 7.29 ng/mL and 9.86 ng/mL, respectively.

A29L Antibody Bioactivity Verification

Using ELISA, the Monkeypox virus A29L antigen (C1619) was used to detect MPXV A29L monoclonal antibody R395a6. The EC50 was 10.54 ng/mL.

B6R Antibody Bioactivity Verification

Using ELISA, the Monkeypox virus B6R antigen (C1622) was used to detect MPXV B6R monoclonal antibody R401c8. The EC50 was 15.26 ng/mL.

A35R Antibody Colloidal Gold Platform Verification

Colloidal gold strips were prepared using the Monkeypox virus A35R antibody. The strips were coated with A35R antibodies R386t7, R387c6, and R384c2, and labeled with A35R antibody R392a3. The A35R antigen (C1620) was detected at concentrations of 0, 0.01, 0.1, 1, 10, and 100 ng/mL. The T line color showed a clear gradient change, with all three antibody pairs having a detection sensitivity of 10 pg/mL.

A29L Antibody Colloidal Gold Platform Verification

Colloidal gold strips were prepared using the Monkeypox virus A29L antibody. The strips were coated with A29L antibody R396t5 and labeled with A29L antibodies R399e2 and R397e8. The A29L antigen (C1619) was detected at concentrations of 0, 0.01, 0.1, 1, 10, and 100 ng/mL. The T line color showed a clear gradient change, with both antibody pairs having a detection sensitivity of 10 pg/mL.

B6R Antibody Colloidal Gold Platform Verification

Colloidal gold strips were prepared using the Monkeypox virus B6R antibody. The strips were coated with B6R antibody R401c8 and labeled with B6R antibody R400e7. The B6R antigen (C1622) was detected at concentrations of 0, 0.01, 0.1, 1, 10, and 100 ng/mL. The T line color showed a clear gradient change, with a detection sensitivity of 10 pg/mL for both antibodies.

M1R Antibody Colloidal Gold Platform Verification

Colloidal gold strips were prepared using the Monkeypox virus M1R antibody. The strips were coated with M1R antibody R417t6 and labeled with M1R antibody R418t3. The M1R antigen (C1624) was detected at concentrations of 0, 0.0125, 0.05, 0.1, and 1 ng/mL. The T line color showed a clear gradient change, with a detection sensitivity of 10 pg/mL for both antibodies.

Monkeypox virus (MPXV) is an enveloped double-stranded DNA virus belonging to the Orthopoxvirus genus. MPXV is approximately 200x250 nm in size, with an ovoid or brick-shaped structure. The core is surrounded by a double-layer lipid membrane, which has randomly arranged tubular elements on its surface. MPXV can effectively infect primary human monocytes. Unlike Cowpox virus (CPV), MPXV does not interfere with MHC expression or intracellular transport to influence immune responses. Instead, it suppresses CD4+ and CD8+ T cell activation, eliminates local T cell responses, and avoids systemic immunosuppression and immune surveillance.
The MPXV genome is a linear double-stranded DNA molecule (about 197 kb) with inverted terminal repeats (ITRs) at the ends. The ITRs contain a set of short tandem repeat sequences and multiple open reading frames (ORFs). The central region of the genome encodes housekeeping genes, while the terminal variable regions encode virulence and host range genes. MPXV has a low dependency on host cell DNA and RNA replication, and it replicates intracellular mature viruses (IMV) and extracellular enveloped viruses (EEV) in the cytoplasm of the host cell. IMVs have a robust physical structure for inter-host transmission, while the more fragile EEVs are enveloped, limiting host immune clearance and facilitating intercellular virus spread.

Reference

1.Sarah Keasey, Christine Pugh, Alexander Tikhonov, et al. "Proteomic Basis of the Antibody Response to Monkeypox Virus Infection Examined in Cynomolgus Macaques and a Comparison to Human Smallpox Vaccination." Plos One, 2010, 5(12): e15547.
2.Emmanuel Alakunle, Ugo Moens, Godwin Nchind, et al. "Monkeypox Virus in Nigeria: Infection Biology, Epidemiology, and Evolution." Viruses, 2020, 12(11): 1257.
3.S. N. Shchelkunov, A. V. Totmenin, P. F. Safronov, et al. "Analysis of the Monkeypox Virus Genome." Virology, 2002, 297: 172-194.