Improved tumor targeting of anti–epidermal growth factor receptor Nanobodies through albumin binding: taking advantage of modular Nanobody technology

Tijink, Bernard M.; Laeremans, Toon; Budde, Marianne; Walsum, Marijke Stigter-van; Dreier, Torsten; Haard, Hans J. de; Leemans, C. René; Dongen, Guus A.M.S. van

doi:10.1158/1535-7163.mct-07-2384

Cited by 267 publications

(216 citation statements)

References 29 publications

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“…Nevertheless, the bsFab format authorizes the linker-free addition of single-domain antibodies at the C-terminus of CH1 or Ck domains (data not shown). It can therefore be envisaged to construct trispecific formats by adding an anti-human serum albumin sdAb, a method that has been shown to significantly increase tumor retention and half-life (50,51).…”

Section: Discussionmentioning

confidence: 99%

Single-Domain Antibody–Based and Linker-Free Bispecific Antibodies Targeting FcγRIII Induce Potent Antitumor Activity without Recruiting Regulatory T Cells

Rozan

Cornillon

Pétiard

et al. 2013

Molecular Cancer Therapeutics

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Antibody-dependent cell-mediated cytotoxicity, one of the most prominent modes of action of antitumor antibodies, suffers from important limitations due to the need for optimal interactions with Fcg receptors. In this work, we report the design of a new bispecific antibody format, compact and linker-free, based on the use of llama single-domain antibodies that are capable of circumventing most of these limitations. This bispecific antibody format was created by fusing single-domain antibodies directed against the carcinoembryonic antigen and the activating FcgRIIIa receptor to human Ck and CH1 immunoglobulin G1 domains, acting as a natural dimerization motif. In vitro and in vivo characterization of these Fab-like bispecific molecules revealed favorable features for further development as a therapeutic molecule. They are easy to produce in Escherichia coli, very stable, and elicit potent lysis of tumor cells by human natural killer cells at picomolar concentrations. Unlike conventional antibodies, they do not engage inhibitory FcgRIIb receptor, do not compete with serum immunoglobulins G for receptor binding, and their cytotoxic activity is independent of Fc glycosylation and FcgRIIIa polymorphism. As opposed to anti-CD3 bispecific antitumor antibodies, they do not engage regulatory T cells as these latter cells do not express FcgRIII. Studies in nonobese diabetic/severe combined immunodeficient gamma mice xenografted with carcinoembryonic antigen-positive tumor cells showed that Fab-like bispecific molecules in the presence of human peripheral blood mononuclear cells significantly slow down tumor growth. This new compact, linker-free bispecific antibody format offers a promising approach for optimizing antibody-based therapies.

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Section: Discussionmentioning

confidence: 99%

Single-Domain Antibody–Based and Linker-Free Bispecific Antibodies Targeting FcγRIII Induce Potent Antitumor Activity without Recruiting Regulatory T Cells

Rozan

Cornillon

Pétiard

et al. 2013

Molecular Cancer Therapeutics

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“…These data indicate that in vitro functional assays are not fully predictive for assessment of the in vivo therapeutic potential of aHGF-targeting ligands. One aspect in favor of Nanobodies in comparison with traditional full-length mAbs is their faster and deeper tumor penetration as was previously observed for the aEGFR-Nanobody formats (18).…”

Section: Mol Cancer Ther; 11(4) April 2012mentioning

confidence: 76%

“…Therefore, monospecific Nanobodies do not seem to be qualified for long-term blockage of growth factors and their receptors. Improvement of the pharmacokinetic and dynamic properties of otherwise short-live molecules can be achieved by fusing an anti-albumin unit (aAlb) to the Nanobody, as described by Tijink and colleagues (18). They compared the biodistribution of a bivalent aEGFRNanobody (aEGFR-aEGFR) with a trivalent Nanobody construct containing the aAlb-unit (aEGFR-aEGFRaAlb) in nude mice bearing A431 tumors.…”

Section: Introductionmentioning

confidence: 99%

Nanobodies Targeting the Hepatocyte Growth Factor: Potential New Drugs for Molecular Cancer Therapy

Vosjan

Vercammen

Kolkman

et al. 2012

Molecular Cancer Therapeutics

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115

114

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Hepatocyte growth factor (HGF) and its receptor c-Met are associated with increased aggressiveness of tumors and poor prognostic outcome of patients with cancer. Here, we report the development and characterization of therapeutic anti-HGF (aHGF)-Nanobodies and their potential for positron emission tomographic (PET) imaging to assess HGF expression in vivo. Two aHGF-Nanobodies designated 1E2 and 6E10 were identified, characterized, and molecularly fused to an albumin-binding Nanobody unit (Alb8) to obtain serum half-life extension. The resulting Nanobody formats were radiolabeled with the positron emitter zirconium-89 ( 89 Zr, t1/ 2 ¼ 78 hours), administered to nude mice bearing U87 MG glioblastoma xenografts, and their biodistribution was assessed. In addition, their therapeutic effect was evaluated in the same animal model at doses of 10, 30, or 100 mg per mouse. The 89 Zr-Nanobodies showed similar biodistribution with selective tumor targeting. For example, 1E2-Alb8 showed decreased blood levels of 12.6%ID/g AE 0.6%ID/g, 7.2%ID/g AE 1.0%ID/g, 3.4%ID/g AE 0.3%ID/g, and 0.3%ID/g AE 0.1%ID/g at 1, 2, 3, and 7 days after injection, whereas tumor uptake levels remained relatively stable at these time points: 7.8%ID/g AE 1.1%ID/g, 8.9%ID/g AE 1.0%ID/g, 8.7%ID/g AE 1.5%ID/g, and 7.2%ID/g AE1.6%ID/g. Uptake in normal tissues was lower than in tumor, except for kidneys. In a therapy study, all Nanobody-treated mice showed tumor growth delay compared with the control saline group. In the 100-mg group, four of six mice were cured after treatment with 1E2-Alb8 and 73 days follow-up, and three of six mice when treated with 6E10-Alb8. These results provide evidence that Nanobodies 1E2-Alb8 and 6E10-Alb8 have potential for therapy and PET imaging of HGFexpressing tumors.

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“…Pour éviter ce problème, une première approche consiste à coupler chimiquement une molécule de PEG (polyéthylène glycol) au dAb (PEGylation) [16]. Une solution bien plus élégante consiste à fusionner un dAb d'intérêt à un autre dAb affin pour l'albumine, protéine très abondante dans le sérum, conférant ainsi à la protéine de fusion un temps de demi-vie bien plus long, dépendant de l'affinité du dAb anti-albumine [17]. Finalement, une autre solution consiste à multimériser les dAb [18], de façon à augmenter simultanément le poids moléculaire et l'affinité apparente de la molécule résultante.…”

Section: Ingénierie Et Application Des Dabunclassified

Fragments d’anticorps à domaine unique

Chames

Baty

2009

Med Sci (Paris)

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Improved tumor targeting of anti–epidermal growth factor receptor Nanobodies through albumin binding: taking advantage of modular Nanobody technology

Abstract: The f15-kDa variable domains of camelid heavy-chainonly antibodies (called Nanobodies) can easily be formatted as multivalent or multispecific single-chain proteins.

Cited by 267 publications

References 29 publications

Single-Domain Antibody–Based and Linker-Free Bispecific Antibodies Targeting FcγRIII Induce Potent Antitumor Activity without Recruiting Regulatory T Cells

Single-Domain Antibody–Based and Linker-Free Bispecific Antibodies Targeting FcγRIII Induce Potent Antitumor Activity without Recruiting Regulatory T Cells

Nanobodies Targeting the Hepatocyte Growth Factor: Potential New Drugs for Molecular Cancer Therapy

Fragments d’anticorps à domaine unique

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