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• W2018658367 abstract "We have previously developed a chemically conjugated anti-rhesus monkey CD3 immunotoxin FN18−CRM9 that can deplete in vivo T cells and induce long term tolerance of mismatched renal allograft in rhesus monkeys. This immunotoxin is a monkey analogue of anti-human CD3 immunotoxin UCHT1−CRM9. In this study, we cloned the light and heavy chain variable regions of anti-monkey CD3 monoclonal antibody FN18 and constructed a single-chain Fv (sFv) by linking variable light and variable heavy regions with a (Gly4Ser)3 linker. The single-chain immunotoxin DT390−FN18sFv was constructed by ligating the sFv to the carboxyl terminus of DT390, a truncated form of diphtheria toxin. The DT390−FN18sFv fusion protein was expressed in Escherichia coli and purified with Ni-RTA affinity and anion exchange columns. Similar to the chemically conjugated immunotoxin FN18−CRM9, DT390−FN18sFv can also specifically inhibit protein synthesis in primary monkey T cells in a dose-dependent manner. DT390−FN18sFv at 10-7 mol/L or FN18−CRM9 at 10-8 mol/L is sufficient to reduce protein synthesis of monkey primary T cells to less than 5% of the control. The 50% inhibition dosage (IC50) of FN18−CRM9 is 1 × 10-10 mol/L, while the IC50 of DT390−FN18sFv is 1 × 10-8 mol/L, reflecting the lowered affinity of monovalent Fab‘ FN18 to its parental divalent antibody. The availability of functional FN18sFv will provide the basis for the construction of divalent anti-CD3 immunotoxins for preclinical studies on the induction of tolerance in organ transplantation and experimental autoimmune diseases." @default.
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• W2018658367 date "1997-09-01" @default.
• W2018658367 modified "2023-09-25" @default.
• W2018658367 title "Genetic Construction and Characterization of an Anti-Monkey CD3 Single-Chain Immunotoxin with a Truncated Diphtheria Toxin" @default.
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• W2018658367 doi "https://doi.org/10.1021/bc9701398" @default.
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