Fig. 4.

Affinity-matured anti-TNF-α antibodies have improved neutralizing abilities. L929 cells were treated with actinomycin and TNF-α, as indicated for 24 h, in the absence and presence of purified anti-TNF-α scFv antibodies. WST-1 was added and incubated for an additional 4 h, and OD₄₅₀ was recorded. Fifty percent neutralizing dose values were 141 pM for D2E7 (open green circle), 3.5 pM for A1 (red square), 6.1 pM for cb1-3 (red triangle), and 6.5 pM for cb2-6 (red diamond). Data represent mean ± SEM with each measurement performed in quadruplicate.