Construction and expression of an anti-VEGFR2 Nanobody-Fc fusionbody in NS0 host cell

Qasemi, Maryam; Behdani, Mahdi; Shokrgozar, Mohammad Ali; Molla-Kazemiha, Vahid; Mohseni-Kuchesfahani, Homa; Habibi‐Anbouhi, Mahdi

doi:10.1016/j.pep.2016.03.004

Cited by 21 publications

(12 citation statements)

References 28 publications

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“…For therapeutic applications, nanobodies can readily be humanized, e.g. by fusion to the hinge and Fc-domains of human IgG1 31 . Moreover, the VHH domain itself can be humanized by substituting framework residues to more closely resemble those of human VH domains 32 .…”

Section: Discussionmentioning

confidence: 99%

Nanobodies effectively modulate the enzymatic activity of CD38 and allow specific imaging of CD38+ tumors in mouse models in vivo

Fumey

Koenigsdorf

Kunick

et al. 2017

Sci Rep

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The cell surface ecto-enzyme CD38 is a promising target antigen for the treatment of hematological malignancies, as illustrated by the recent approval of daratumumab for the treatment of multiple myeloma. Our aim was to evaluate the potential of CD38-specific nanobodies as novel diagnostics for hematological malignancies. We successfully identified 22 CD38-specific nanobody families using phage display technology from immunized llamas. Crossblockade analyses and in-tandem epitope binning revealed that the nanobodies recognize three different non-overlapping epitopes, with four nanobody families binding complementary to daratumumab. Three nanobody families inhibit the enzymatic activity of CD38 in vitro, while two others were found to act as enhancers. In vivo, fluorochrome-conjugated CD38 nanobodies efficiently reach CD38 expressing tumors in a rodent model within 2 hours after intravenous injection, thereby allowing for convenient same day in vivo tumor imaging. These nanobodies represent highly specific tools for modulating the enzymatic activity of CD38 and for diagnostic monitoring CD38-expressing tumors.

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Section: Discussionmentioning

confidence: 99%

Nanobodies effectively modulate the enzymatic activity of CD38 and allow specific imaging of CD38+ tumors in mouse models in vivo

Fumey

Koenigsdorf

Kunick

et al. 2017

Sci Rep

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“…Rituximab (anti-CD20) and cetuximab (anti-EGFR), for example, each carry the VH and VL domains of its parental mouse mAb fused to the constant domains of the human IgG1 heavy chain and the human kappa light chain. Analogous chimeric nanobody-human IgG heavy chain antibodies can be generated by fusion of the VHH encoding region to the hinge and Fc domains of a human IgG ( 37 – 39 ).…”

Section: Conventional and Heavy Chain Antibodies As Antitumor Therapementioning

confidence: 99%

Nanobodies and Nanobody-Based Human Heavy Chain Antibodies As Antitumor Therapeutics

Bannas¹,

2017

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Monoclonal antibodies have revolutionized cancer therapy. However, delivery to tumor cells in vivo is hampered by the large size (150 kDa) of conventional antibodies. The minimal target recognition module of a conventional antibody is composed of two non-covalently associated variable domains (VH and VL). The proper orientation of these domains is mediated by their hydrophobic interface and is stabilized by their linkage to disulfide-linked constant domains (CH1 and CL). VH and VL domains can be fused via a genetic linker into a single-chain variable fragment (scFv). scFv modules in turn can be fused to one another, e.g., to generate a bispecific T-cell engager, or they can be fused in various orientations to antibody hinge and Fc domains to generate bi- and multispecific antibodies. However, the inherent hydrophobic interaction of VH and VL domains limits the stability and solubility of engineered antibodies, often causing aggregation and/or mispairing of V-domains. Nanobodies (15 kDa) and nanobody-based human heavy chain antibodies (75 kDa) can overcome these limitations. Camelids naturally produce antibodies composed only of heavy chains in which the target recognition module is composed of a single variable domain (VHH or Nb). Advantageous features of nanobodies include their small size, high solubility, high stability, and excellent tissue penetration in vivo. Nanobodies can readily be linked genetically to Fc-domains, other nanobodies, peptide tags, or toxins and can be conjugated chemically at a specific site to drugs, radionuclides, photosensitizers, and nanoparticles. These properties make them particularly suited for specific and efficient targeting of tumors in vivo. Chimeric nanobody-heavy chain antibodies combine advantageous features of nanobodies and human Fc domains in about half the size of a conventional antibody. In this review, we discuss recent developments and perspectives for applications of nanobodies and nanobody-based human heavy chain antibodies as antitumor therapeutics.

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“…In addition the VHHs are interesting alternative to the use of monoclonal antibodies for therapy as shown by recent studies of their use against the Dengue virus [19], H5N1 influenza [20], viral infection [21], aflatoxins in agro-products [22], head and neck cancers [23], vascular endothelial growth factor implicated in cancers [24], venom therapy [25] and Plasmodium knowlesi malaria vector [26]. Phase II clinical trials are currently underway to evaluate the efficacy and toxicity of a therapeutic Nanobody TM in patients with thrombotic thrombocytopenic purpura [27] showing recent promising results [28].…”

Section: Introductionmentioning

confidence: 99%

Global analysis of VHHs framework regions with a structural alphabet

Noël

Malpertuy²,

Brevern

2016

Biochimie

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The VHHs are antigen-binding region/domain of camelid heavy chain antibodies (HCAb). They have many interesting biotechnological and biomedical properties due to their small size, high solubility and stability, and high affinity and specificity for their antigens. HCAb and classical IgGs are evolutionary related and share a common fold. VHHs are composed of regions considered as constant, called the frameworks (FRs) connected by Complementarity Determining Regions (CDRs), a highly variable region that provide interaction with the epitope. Actually, no systematic structural analyses had been performed on VHH structures despite a significant number of structures. This work is the first study to analyse the structural diversity of FRs of VHHs. Using a structural alphabet that allows approximating the local conformation, we show that each of the four FRs do not have a unique structure but exhibit many structural variant patterns. Moreover, no direct simple link between the local conformational change and amino acid composition can be detected. These results indicate that long-range interactions affect the local conformation of FRs and impact the building of structural models.

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Construction and expression of an anti-VEGFR2 Nanobody-Fc fusionbody in NS0 host cell

Cited by 21 publications

References 28 publications

Nanobodies effectively modulate the enzymatic activity of CD38 and allow specific imaging of CD38+ tumors in mouse models in vivo

Nanobodies effectively modulate the enzymatic activity of CD38 and allow specific imaging of CD38+ tumors in mouse models in vivo

Nanobodies and Nanobody-Based Human Heavy Chain Antibodies As Antitumor Therapeutics

Global analysis of VHHs framework regions with a structural alphabet

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