Humanization of the murine anti-human CD3 monoclonal antibody OKT3

Humanization of the Bispecific Epidermal Growth Factor Receptor × CD3 Diabody and Its Efficacy as a Potential Clinical Reagent

“…Additionally, hEx3 showed the same or slightly higher cytotoxicity than Ex3, despite a 40-fold lower affinity for EGFR (K a values of the mouse and humanized 528 Fv to EGFR, as estimated from isothermal titration calorimetry, are 8.17 Â 10 8 and 1.89 Â 10 7 , respectively). K a values against to CD3 of the diabody consisted of mouse OKT3 is 4.5 Â 10 7 for reference, and it is reported that mouse and humanized OKT3 show almost same property (34,39). hEx3 did not enhance T-LAK cytotoxicity against MCF7 or non -EGFR-transfected CHO cells (Fig.…”

“…The genes for the humanized anti-CD3 antibody OKT3 Fv were similarly constructed with whole synthesized and E. coli codon-optimized genes (31 -33). The VH and VL regions of humanized 528 Fv are designated h5H and h5L, and those of humanized OKT3 Fv, used a previously published humanized sequence, are designated hOH and hOL, respectively (34). The two hetero-scFvs of humanized anti-EGFR Â anti-CD3 bispecific diabody (designated hEx3) are designated h5HhOL and hOHh5L, and each corresponding gene (h5HhOL and hOHh5L) was inserted into the pRA vector, which is a previously constructed T7 promoter -based expression vector ( Fig.…”

Human derived dimerization tag enhances tumor killing potency of a T-cell engaging bispecific antibody

“…The sequences of anti-GD2 antibody 5F11 20 and humanized anti-CD3 38 have previously been reported. 5F11 scFv (VH-VL orientation) and anti-CD3 hOKT3 scFv (VH-VL) with an orientation VHVL were genetically assembled by a 15 amino acid linker ((GGGGS)5) and synthesized separately (GenScript, Piscataway, NJ).…”

Characterization of a surrogate murine antibody to model anti-human CD3 therapies

“…This dose was selected based on previously published efficacy results 56 and on the safety profile of 145-2C11, which induces signs of severe toxicity at higher doses. 2C11-Novi was administered at the same dose as 145-2C11 and the higher dose To alter the risk-to-benefit ratio and thus allow assessment of CD3-directed therapies for the treatment of autoimmune diseases, humanized versions of rodent anti-human CD3ε mAbs [22][23][24][25][26][27] and a fully human mAb 28 were created. These mAbs have a similar engineered element built into their Fc portion, i.e., mutations introduced into the second heavy constant domain to reduce FcγR binding and cytokine release associated with an Fc-dependent mechanism.…”