Generation and characterization of rendomab-B1, a monoclonal antibody displaying potent and specific antagonism of the human endothelin B receptor

Allard, Bertrand; Wijkhuisen, Anne; Borrull, Aurélie; Deshayes, Frédérique; Priam, Fabienne; Lamourette, Patricia; Ducancel, Frédéric; Boquet, Didier; Couraud, Jean-Yves

doi:10.4161/mabs.22696

Cited by 20 publications

(27 citation statements)

References 53 publications

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“…1C shows their apparent affinities for UACC-257, and a detailed analysis of rendomab-B4 binding on the 3 different melanoma cells is shown below. In addition, among the 23 new antibodies that we tested, some other mAbs were totally unable to bind ETB expressed on any tumoral cell line, as was observed for the rendomab B1 that we already described, 25 whereas others recognized ETB expressed only on a given tumoral cell line (results not shown). Since, as illustrated in Fig.…”

Section: Characterization Of Anti-human Etb Specific Monoclonal Antibsupporting

confidence: 54%

“…25 However this mAb displayed a very low affinity for ETB expressed on tumor cells, which prompted us to look for the potential presence, among the 23 other anti-ETB mAbs that we produced, of antibodies that could recognize the tumor-associated form of human ETB. Toward that end, the binding properties of all the mAbs on Chinese hamster ovary (CHO) cells stably transfected with human ETB (CHO-ETB) and different melanoma cell lines were investigated.…”

Section: Characterization Of Anti-human Etb Specific Monoclonal Antibmentioning

confidence: 99%

“…Our group 25 and others 17 have recently developed mAbs directed against ETB. However, the higher affinity of the antibody that we describe here, associated with a fast internalization of ETB, might make it a good candidate for antibody-drug conjugate (ADC) development to target ETB in melanoma.…”

Section: Introductionmentioning

confidence: 99%

“…However, the higher affinity of the antibody that we describe here, associated with a fast internalization of ETB, might make it a good candidate for antibody-drug conjugate (ADC) development to target ETB in melanoma. 17 Previously, our group described rendomab-B1, 25 a mAb that specifically recognizes human ETB. This antibody is a strong antagonist and inhibits ETB functions in endothelial cells.…”

Section: Introductionmentioning

confidence: 99%

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Rendomab B4, a monoclonal antibody that discriminates the human endothelin B receptor of melanoma cells and inhibits their migration

Borrull

Allard

Wijkhuisen

et al. 2016

mAbs

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Metastatic melanoma is an aggressive cancer with a poor prognostic, and the design of new targeted drugs to treat melanoma is a therapeutic challenge. A promising approach is to produce monoclonal antibodies (mAbs) against the endothelin B receptor (ETB), which is known to be overexpressed in melanoma and to contribute to proliferation, migration and vasculogenic mimicry associated with invasiveness of this cancer. We previously described rendomab-B1, a mAb produced by DNA immunization. It is endowed with remarkable characteristics in term of affinity, specificity and antagonist properties against human ETB expressed by the endothelial cells, but, surprisingly, had poor affinity for ETB expressed by melanoma cells. This characteristic strongly suggested the existence of a tumor-specific ETB form. In the study reported here, we identified a new mAb, rendomab-B4, which, in contrast to rendomab-B1, binds ETB expressed on UACC-257, WM-266-4 and SLM8 melanoma cells. Moreover, after binding to UACC-257 cells, rendomab-B4 is internalized and colocalizes with the endosomal protein EEA-1. Interestingly, rendomab-B4, despite its inability to compete with endothelin binding, is able to inhibit phospholipase C pathway and migration induced by endothelin. By contrast, rendomab-B4 fails to decrease ERK1/2 phosphorylation induced by endothelin, suggesting a biased effect on ETB. These particular properties make rendomab-B4 an interesting tool to analyze ETB-structure/function and a promising starting point for the development of new immunological tools in the field of melanoma therapeutics.

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Section: Characterization Of Anti-human Etb Specific Monoclonal Antibsupporting

confidence: 54%

Section: Characterization Of Anti-human Etb Specific Monoclonal Antibmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Rendomab B4, a monoclonal antibody that discriminates the human endothelin B receptor of melanoma cells and inhibits their migration

Borrull

Allard

Wijkhuisen

et al. 2016

mAbs

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“…[38][39][40] Genetic immunization, where antigen is produced within the host, thus overcoming the requirement for purified protein or cells, has also resulted in the generation of functionally active GPCR antibodies. [41][42][43] In the study reported here, we applied in vivo and in vitro methods to generate human monoclonal antibodies to the human CXCR2 receptor and assess whether use of these different approaches can result in the isolation of antibodies that are diverse in both epitope and function. Cell lines overexpressing human CXCR2 have been used as the antigen for whole cell mouse immunization.…”

Section: Introductionmentioning

confidence: 99%

Phage display and hybridoma generation of antibodies to human CXCR2 yields antibodies with distinct mechanisms and epitopes

Rossant

Carroll²,

Huang

et al. 2014

mAbs

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Refolding of Aggregation-Prone ScFv Antibody Fragments Assisted by Hydrophobically Modified Poly(sodium acrylate) Derivatives

et al. 2016

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ScFv antibody fragments are a promising alternatives to full-length antibodies for both therapeutic and diagnosis applications. They can be overexpressed in bacteria, which enables easy large scale production. Since scFv are artificial constructs, they are poorly soluble and prone to aggregation, which makes them difficult to manipulate and to refold. Here, we report stabilization and refolding of scFv fragments from urea-unfolded solutions based on the use of micromolar amounts of polymers playing the role of artificial chaperons. Using fluorescence correlation spectroscopy, we determined the size and aggregation number of complexes of scFv with unmodified or hydrophobically-modified poly(sodium acrylate). The evolution of the secondary structure along the refolding procedure, in the presence or absence of 0.4 M L-arginine at scFv:polymer < 1:5 wt/wt, was determined by high-sensitivity synchrotron-radiation circular dichroism, SRCD. Measurements revealed that refolding in the presence of polymers yields native-like secondary structure, though a different folding pathway can be followed compared to refolding in the absence of polymer. This is the first report on the use of macromolecular additives to assist refolding of a multi-domain protein of therapeutic interest.3

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Generation and characterization of rendomab-B1, a monoclonal antibody displaying potent and specific antagonism of the human endothelin B receptor

Abstract: Generation and characterization of rendomab-B1, a monoclonal antibody displaying potent and specific antagonism of the human endothelin B receptor, mAbs, 5:1, 56-69,

Cited by 20 publications

References 53 publications

Rendomab B4, a monoclonal antibody that discriminates the human endothelin B receptor of melanoma cells and inhibits their migration

Rendomab B4, a monoclonal antibody that discriminates the human endothelin B receptor of melanoma cells and inhibits their migration

Phage display and hybridoma generation of antibodies to human CXCR2 yields antibodies with distinct mechanisms and epitopes

Refolding of Aggregation-Prone ScFv Antibody Fragments Assisted by Hydrophobically Modified Poly(sodium acrylate) Derivatives

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