Recombinant Mouse Anti-VACV Protein L1 Antibody (M12B9)

(A) Seven anti-A27 MAbs were tested for their ability to bind purified A2716–100 protein. All MAbs except MAb 8E3 successfully bound the construct. Anti-D8 MAb Ab12.1 (aD8 Ab12.1) was used as a negative control. (B) Direct comparison of four selected anti-A27 MAbs binding the A2716–100 protein versus the A2716–110 protein. All four tested anti-A27 MAbs, including MAb 8E3, bound the longer construct. Anti-L1 MAb M12B9 (aL1 MAb M12B9) was used as a negative control. Dashed lines, the cutoffs for positive results (OD, 1). The experiments were performed three times.

Kaever, T. , Matho, M. H. , Meng, X. , Crickard, L. , Schlossman, A. , & Xiang, Y. , et al. (2016). Linear epitopes in vaccinia virus a27 are targets of protective antibodies induced by vaccination against smallpox. Journal of Virology, 90(9), 4334.

Anti-A27 antibodies were tested for their ability to neutralize in vitro in a fluorescence-activated cell sorting-based neutralization assay featuring MAbs 1G6, 12G2, 8H10, 6F11, 4G5, 12C3, and 8E3. Anti-L1 MAb M12B9 was used as a positive control. All antibodies were used at a final concentration of 20 μg/ml. Anti-A27 MAbs 1G6, 12G2, and 8H10 (group I) were capable of strong neutralization (>90%) in the presence of complement (bottom) but not in its absence (top). Antibodies 6F11 (group II), 4B5 and 12C3 (group III), and 8E3 (group IV) did not neutralize in the absence of complement and showed weak or no neutralization ability (<20%) in the presence of complement. The positive control, anti-L1 MAb M12B9, strongly neutralized (>95%) the virus in a complement-independent manner. Dashed lines, 50% neutralization. The experiments were performed at least twice.

Kaever, T. , Matho, M. H. , Meng, X. , Crickard, L. , Schlossman, A. , & Xiang, Y. , et al. (2016). Linear epitopes in vaccinia virus a27 are targets of protective antibodies induced by vaccination against smallpox. Journal of Virology, 90(9), 4334.