Therapeutic antibody fragments with prolonged in vivo half-lives

Chapman, Andrew P.; Antoniw, Pari; Spitali, Mariangela; West, Shauna; Stephens, Sue; King, David J.

doi:10.1038/11717

Cited by 325 publications

(191 citation statements)

References 12 publications

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“…In addition, the plasma concentrations of the Fab’PEG format were sustained for longer (Figure 4); this may be due not only to the higher molar dose, but also due to the presence of a PEG moiety. The PEG moiety is responsible for extending half-life in an FcRn-independent manner, 46 and therefore could provide a PK advantage compared with the IgG formats that are blocking their own recycling mechanism. Rozanolixizumab and the triFab’ format were dosed approximately equally in molar terms, but the IgG4P format showed greater activity (Figure 3A), although this might, in part, be due to prolonged PK of rozanolixizumab (Figure 4).…”

Section: Resultsmentioning

confidence: 99%

“…Humanized Fab’ was expressed in, and prepared from E. coli , 46,70 and further modified as detailed in Supplementary Table 3. IgG molecules were prepared from CHO SXE 69 cell culture medium by affinity chromatography on MabSelect™ SuRe™ (GE Healthcare, UK), followed by size-exclusion chromatography (SEC) on HiLoad XK 50/60 Superdex 200 PG (GE Healthcare, UK).…”

Section: Methodsmentioning

confidence: 99%

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Generation and characterization of a high affinity anti-human FcRn antibody, rozanolixizumab, and the effects of different molecular formats on the reduction of plasma IgG concentration

Smith

Kießling

Lledó-García

et al. 2018

mAbs

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Rozanolixizumab (UCB7665), a humanized high-affinity anti-human neonatal Fc receptor (FcRn) monoclonal antibody (IgG4P), has been developed to reduce pathogenic IgG in autoimmune and alloimmune diseases. We document the antibody isolation and compare rozanolixizumab with the same variable region expressed in various mono-, bi- and trivalent formats. We report activity data for rozanolixizumab and the different molecular formats in human cells, FcRn-transgenic mice, and cynomolgus monkeys. Rozanolixizumab, considered the most effective molecular format, dose-dependently and selectively reduced plasma IgG concentrations in an FcRn-transgenic mouse model (no effect on albumin). Intravenous (IV) rozanolixizumab dosing in cynomolgus monkeys demonstrated non-linear pharmacokinetics indicative of target-mediated drug disposition; single IV rozanolixizumab doses (30 mg/kg) in cynomolgus monkeys reduced plasma IgG concentration by 69% by Day 7 post-administration. Daily IV administration of rozanolixizumab (initial 30 mg/kg loading dose; 5 mg/kg daily thereafter) reduced plasma IgG concentrations in all cynomolgus monkeys, with low concentrations maintained throughout the treatment period (42 days). In a 13-week toxicology study in cynomolgus monkeys, supra-pharmacological subcutaneous and IV doses of rozanolixizumab (≤ 150 mg/kg every 3 days) were well tolerated, inducing sustained (but reversible) reductions in IgG concentrations by up to 85%, with no adverse events observed. We have demonstrated accelerated natural catabolism of IgG through inhibition of IgG:FcRn interactions in mice and cynomolgus monkeys. Inhibition of FcRn with rozanolixizumab may provide a novel therapeutic approach to reduce pathogenic IgG in human autoimmune disease. Rozanolixizumab is being investigated in patients with immune thrombocytopenia (NCT02718716) and myasthenia gravis (NCT03052751).

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Generation and characterization of a high affinity anti-human FcRn antibody, rozanolixizumab, and the effects of different molecular formats on the reduction of plasma IgG concentration

Smith

Kießling

Lledó-García

et al. 2018

mAbs

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“…Interestingly, the P-SMAC showed significantly improved circulating half-life (∼5-6 h) compared with the unconjugated Fab (∼1 h), perhaps because of the increased overall hydrophobicity of the Fab after conjugation (Fig. S3) (38,39). Of note, P-SMAC has an improved serum half-life relative to small bispecific scFvs such as bispecific T-cell engagers, which have a half-life of ∼2 h in humans, despite their similar molecular weight (∼50,000 Da) (40,41).…”

Section: Resultsmentioning

confidence: 99%

Bispecific small molecule–antibody conjugate targeting prostate cancer

Kim

Axup

Lawson

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Bispecific antibodies, which simultaneously target CD3 on T cells and tumor-associated antigens to recruit cytotoxic T cells to cancer cells, are a promising new approach to the treatment of hormone-refractory prostate cancer. Here we report a site-specific, semisynthetic method for the production of bispecific antibodylike therapeutics in which a derivative of the prostate-specific membrane antigen-binding small molecule DUPA was selectively conjugated to a mutant αCD3 Fab containing the unnatural amino acid, p-acetylphenylalanine, at a defined site. Homogeneous conjugates were generated in excellent yields and had good solubility. The efficacy of the conjugate was optimized by modifying the linker structure, relative binding orientation, and stoichiometry of the ligand. The optimized conjugate showed potent and selective in vitro activity (EC 50 ∼100 pM), good serum half-life, and potent in vivo activity in prophylactic and treatment xenograft mouse models. This semisynthetic approach is likely to be applicable to the generation of additional bispecific agents using drug-like ligands selective for other cell-surface receptors.antibody engineering | immunotherapy

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“…Monoclonal antibodies (MAbs) are widely used for a range of biological assays and clinical therapies (Chapman et al 1999). The production of large amounts of MAbs at low cost is required for industrial application (Fussenegger et al 1999).…”

Section: Introductionmentioning

confidence: 99%

Growth control of hybridoma cells with an artificially induced EpoR-gp130 heterodimer

et al. 2006

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IL-6 has been known to modulate the growth of many hybridoma cells and also promote resultant antibody productivity. However, IL-6 is so expensive that the use of IL-6-dependent hybridomas for industrial antibody production is not practical. In this study, we aimed at designing antibody/gp130 and antibody/EpoR chimeras which could tightly control cell growth in response to more affordable cognate antigen. Retroviral vectors encoding V H or V L region of anti-hen egg lysozyme (HEL) antibody HyHEL-10 tethered to a pair of extracellular D2/transmembrane domains of erythropoietin receptor (EpoR) and cytoplasmic domains of either EpoR or gp130, were constructed, and a homodimeric or a heterodimeric pair of chimeric receptor combinations (V H -gp130 and V L -gp130 or V H -gp130 and V L -EpoR) were expressed in an IL-6-dependent hybridoma 7TD1. The chimeric receptor-derived growth signal was observed in both combinations, while some residual growth signal was observed in the absence of HEL. To reduce interchain interaction between the two receptor chains, we introduced mutations to the transmembrane domain of both chimera combinations. Consequently, the heterodimeric combination of V H -gp130 and V L -EpoR showed clear HEL-dependent cell growth, while the homodimeric combination of V H -gp130 and V L -gp130 showed reduced cell growth in the absence of HEL. This is the first report that an EpoR-gp130 cytoplasmic domain heterodimer could transduce a growth signal in hybridoma cells, indicating tight and economical growth control of hybridoma cells via our chimeric receptors.

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Therapeutic antibody fragments with prolonged in vivo half-lives

Cited by 325 publications

References 12 publications

Generation and characterization of a high affinity anti-human FcRn antibody, rozanolixizumab, and the effects of different molecular formats on the reduction of plasma IgG concentration

Generation and characterization of a high affinity anti-human FcRn antibody, rozanolixizumab, and the effects of different molecular formats on the reduction of plasma IgG concentration

Bispecific small molecule–antibody conjugate targeting prostate cancer

Growth control of hybridoma cells with an artificially induced EpoR-gp130 heterodimer

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