Recombinant Llama Anti-MARV NP Single Domain Antibody (PABC-561) (CAT#: PABC-561)

Figure 1 Titrations of fusions of nluc–NP C-terminal domains of either MARV or MLAV over oriented sdAb A, B or C.

Sherwood, L. J., & Hayhurst, A. (2019). Periplasmic sdAb-APEX2 Fusions Enable Facile Visualization of Ebola, Marburg, and Mĕnglà virus Nucleoproteins, Alluding to Similar Antigenic Landscapes among Marburgvirus and Dianlovirus. Viruses, 11(4), 364.

Figure 2 ELISA titration of the anti-MARV sdAb monomers and dimers over MARV NP with the highest concentration also applied to Bundibugyo NP.

Bundibugyo NP was expressed at higher levels than Zaire NP and so was convenient to use for controls yet shares high homology at the C-terminal domain for our studies.

Darling, T. L., Sherwood, L. J., & Hayhurst, A. (2017). Intracellular crosslinking of filoviral nucleoproteins with xintrabodies restricts viral packaging. Frontiers in immunology, 8, 1197.

Figure 3 Locating the region of nucleoprotein (NP) bound by sdAb and establishing EC50 values.

Purified recombinant MARV or Bundibugyo (BEBOV) NP polymers were titrated over passively immobilized anti-MARV NP-sdAb, and captured NP subsequently detected with a constant amount of each phage displayed sdAb followed by anti-M13-horseradish peroxidase (HRP) conjugate.

Garza, J. A., Taylor, A. B., Sherwood, L. J., Hart, P. J., & Hayhurst, A. (2017). Unveiling a drift resistant cryptotope within marburgvirus nucleoprotein recognized by llama single-domain antibodies. Frontiers in immunology, 8, 1234.

Figure 4 Locating the region of nucleoprotein (NP) bound by sdAb and establishing EC50 values.

1,000 ng (1), 100 ng (2), or 10 ng (3) of purified MARV NP C-terminus was probed with 100 nM of each sdAb-AP fusion protein for equivalent times and developed side-by-side.

Garza, J. A., Taylor, A. B., Sherwood, L. J., Hart, P. J., & Hayhurst, A. (2017). Unveiling a drift resistant cryptotope within marburgvirus nucleoprotein recognized by llama single-domain antibodies. Frontiers in immunology, 8, 1234.

Figure 5 Locating the region of nucleoprotein (NP) bound by sdAb and establishing EC50 values.

The sdAb-gluc fusions were titrated over passively immobilized mbp-NP600 fusion protein to determine the monovalent EC50 values.

Garza, J. A., Taylor, A. B., Sherwood, L. J., Hart, P. J., & Hayhurst, A. (2017). Unveiling a drift resistant cryptotope within marburgvirus nucleoprotein recognized by llama single-domain antibodies. Frontiers in immunology, 8, 1234.

Figure 6 Locating the region of nucleoprotein (NP) bound by sdAb and establishing EC50 values.

The nluc-NP600 fusions were titrated over oriented immobilized sdAb to determine the EC50 values in a reversed orientation.

Garza, J. A., Taylor, A. B., Sherwood, L. J., Hart, P. J., & Hayhurst, A. (2017). Unveiling a drift resistant cryptotope within marburgvirus nucleoprotein recognized by llama single-domain antibodies. Frontiers in immunology, 8, 1234.

Figure 1 Recombinant Llama Anti-MARV NP Single Domain Antibody (B) (PABC-561) in SDS-PAGE

SDS-PAGE analysis of PABC-561 in reduced (Lane 1) and non-reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result, different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.

The purity of PABC-561 was greater than 95% as determined by SDS-PAGE.

Figure 2 Recombinant Llama Anti-MARV NP Single Domain Antibody (B) (PABC-561) in HPLC

The purity of PABC-561 was greater than 95% as determined by SEC-HPLC.