Table 2.

Biolayer-interferometry of MRU44–4-A1 in different antibody formats and IC₅₀ determined in alamarBlue neutralization with different antibody formats.

Setup	Antibody format	KD [nM]	IC50 [nM]
α-LTX + 2 mM EDTA	MRU44–4-A1 scFv-Fc	8.4	n.a.
	MRU44–4-A1 IgG	6.3	n.a.
	MRU44–4-A1 Fab	1.8	n.a.
α-LTX + 10 mM CaCl2	MRU44–4-A1 scFv-Fc	12.2	0.56
	MRU44–4-A1 IgG	4.2	0.35
	MRU44–4-A1 Fab	2.7	0.34

K_D of MRU44–4-A1 to α-LTX was tested in different antibody formats (scFv-Fc, IgG, Fab) and on monomeric/dimeric and tetrameric α-LTX with BLI, by supplementing buffer with either 2 mM EDTA or 10 mM CaCl₂, respectively. IC₅₀ values of different MRU44–4-A1 antibody formats were determined in alamarBlue neutralization assay. IC₅₀ were not applicable (n.a.) for 2 mM EDTA, as toxin would not be active and thereby have no pathogenic effects on cells and consequently no IC₅₀ of antibodies determinable. Resulting values were determined by using Origin Hill1 non-linear fit.