Effector Cell Recruitment with Novel Fv-based Dual-affinity Re-targeting Protein Leads to Potent Tumor Cytolysis and in Vivo B-cell Depletion

Johnson, Syd; Burke, Stephen P.; Huang, Ling; Gorlatov, Sergey; Li, Hua; Wang, Weili; Zhang, Wenjun; Tuaillon, Nadine; Rainey, Jonah; Barat, Bhaswati; Yang, Yinhua; Jin, Linda X.; Ciccarone, Valentina; Moore, Paul A.; Koenig, Scott; Bonvini, Ezio

doi:10.1016/j.jmb.2010.04.001

Cited by 171 publications

(138 citation statements)

References 49 publications

(61 reference statements)

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“…One is the Diabody (Db), which is a heterodimer composed of 2 polypeptide chains encoding V L A-V H B and V H A-V L B in the order of V H -V L or V L -V H with a linker of 5 amino acids. 23 Its derivatives include, but are not limited to, dsDb (interchain disulfide bond between V L and V H of the same antibody), 24 DART (dual-affinity re-targeting, interchain disulfide bond between 2 V L ), 25 scDb (single chain Diabody), 26 and tandAbs (Diabody dimer via flexible linkers in between). 27 The other is the BiTE Ò (bispecific T-cell engager), which is composed of 2 scFvs connected in tandem by an adjustable linker.…”

Section: Introductionmentioning

confidence: 99%

Production of bispecific antibodies in “knobs-into-holes” using a cell-free expression system

Lee

Tran

et al. 2015

mAbs

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(2015) Production of bispecific antibodies in "knobs-into-holes" using a cell-free expression system , mAbs, 7:1, 231-242, DOI: 10.4161/19420862.2015.989013 To link to this article: https://doi.org/10. 4161/19420862.2015 Bispecific antibodies have emerged in recent years as a promising field of research for therapies in oncology, inflammable diseases, and infectious diseases. Their capability of dual target recognition allows for novel therapeutic hypothesis to be tested, where traditional mono-specific antibodies would lack the needed mode of target engagement. Among extremely diverse architectures of bispecific antibodies, knobs-into-holes (KIHs) technology, which involves engineering C H 3 domains to create either a "knob" or a "hole" in each heavy chain to promote heterodimerization, has been widely applied. Here, we describe the use of a cell-free expression system (Xpress CF) to produce KIH bispecific antibodies in multiple scaffolds, including 2-armed heterodimeric scFv-KIH and one-armed asymmetric BiTE-KIH with tandem scFv. Efficient KIH production can be achieved by manipulating the plasmid ratio between knob and hole, and further improved by addition of prefabricated knob or hole. These studies demonstrate the versatility of Xpress CF in KIH production and provide valuable insights into KIH construct design for better assembly and expression titer.

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Section: Introductionmentioning

confidence: 99%

Production of bispecific antibodies in “knobs-into-holes” using a cell-free expression system

Lee

Tran

et al. 2015

mAbs

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“…CD33 is also expressed by normal myeloid progenitors and treatment with anti CD33 is limited by prolonged cytopenia caused by attrition to the common myeloid progenitors (12). In a recent publication Chichili and colleagues (17) describe the preclinical development and validation of a bispecific antibody [Dual-Affinity Re-Targeting (DART)] which binds to the interleukin-3 receptor CD 123 on the surface of AML blasts and uses anti CD3 to capture cytotoxic T cells and bring them into contact with leukemia (19). Several criteria will determine the clinical success of this approach: (I) the design of the bispecific antibody which affects binding to the target and the effector cell, its in vivo distribution and fate; (II) the quality of the antigen target; (III) adverse side effects from off-target effectsin particular damage to other myeloid tissues and cytokine release syndromes (CRSs) from activated T cells.…”

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confidence: 99%

Antibody darts on target for acute myelogenous leukemia

Barrett

2017

Ann. Transl. Med.

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“…Moreover, Macrogenics reported that they are developing a Dual-Affinity ReTargeting (DART) drug that cross-links CD32b with CD79, a signaling molecule for the BCR (24). Discerning how humoral immune responses can be modulated by cotargeting these receptors is critically important for delineating the effect of this approach in settings of autoimmunity.…”

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confidence: 99%

CD19 and CD32b Differentially Regulate Human B Cell Responsiveness

Karnell¹,

Dimasi²,

Karnell³

et al. 2014

The Journal of Immunology

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B cell activation is regulated by a variety of signals. CD19 positively regulates B cell activation, augmenting signals delivered through the BCR complex. In contrast, CD32b contains an ITIM and negatively regulates BCR signaling. Importantly, there are drugs currently in clinical trials and preclinical development that cross-link CD32b to molecules within the BCR complex. We wanted to address how single engagement versus cotargeting these molecules affects human B cell function. When B cells from healthy individuals were activated by signals that mimic a T cell response (IL-21 costimulation), ligation of CD32b, but not CD19, inhibited B cell expansion and plasma cell (PC) differentiation. In contrast, when B cells were activated through TLR, anti-CD19, but not anti-CD32b, blunted the response. However, when both CD19 and CD32b were coengaged by a bispecific anti-CD19×CD32b Ab, both types of stimuli were potently inhibited. Cross-linking CD19 with CD32b also inhibited Ab-independent functions of B cells, such as HLA upregulation, cytokine production, and the ability of B cells to prime CD4+ T cells. Finally, although cross-linking CD19 and CD32b inhibited PC differentiation of primary B cells, it did not alter Ig production from pre-established PCs. These data elucidate the mechanism by which a complex set of signals determines the fate of B cell responsiveness. Although signals through CD19 influence TLR-driven activation, CD32b impacts the magnitude of the response following IL-21 costimulation. Therefore, simultaneous targeting of multiple surface molecules may be a necessary approach to comprehensively modulate B cell activation in vivo.

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Effector Cell Recruitment with Novel Fv-based Dual-affinity Re-targeting Protein Leads to Potent Tumor Cytolysis and in Vivo B-cell Depletion

Cited by 171 publications

References 49 publications

Production of bispecific antibodies in “knobs-into-holes” using a cell-free expression system

Production of bispecific antibodies in “knobs-into-holes” using a cell-free expression system

Antibody darts on target for acute myelogenous leukemia

CD19 and CD32b Differentially Regulate Human B Cell Responsiveness

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