Site-specific antibody-drug conjugation through an engineered glycotransferase and a chemically reactive sugar

Zhu, Zhongyu; Ramakrishnan, B.; Li, Jinyu; Wang, Yanping; Yang, Feng; Prabakaran, Ponraj; Colantonio, Simona; Dyba, Marcin; Qasba, Pradman K.; Dimitrov, Dimiter S.

doi:10.4161/mabs.29889

Cited by 90 publications

(74 citation statements)

References 37 publications

(49 reference statements)

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“…The purified CHO cell expressed m276 IgG1 was directly used for glycan-based site-specific modification and conjugation as previously described (Zhu et al, 2014) with the following modifications. C2-Azide-Galacotose was used as substrate for the Fc-glycan modification.…”

Section: Star Methodsmentioning

confidence: 99%

Eradication of Tumors through Simultaneous Ablation of CD276/B7-H3-Positive Tumor Cells and Tumor Vasculature

Seaman

Zhu²,

Saha³

et al. 2017

Cancer Cell

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SUMMARY Targeting the tumor vasculature with antibody-drug conjugates (ADCs) is a promising anti-cancer strategy that, in order to be realized, must overcome several obstacles, including identification of suitable targets and optimal warheads. Here, we demonstrate that the cell surface protein CD276/B7-H3 is broadly overexpressed by multiple tumor types on both cancer cells and tumor-infiltrating blood vessels, making it a potentially ideal dual-compartment therapeutic target. In preclinical studies CD276-ADCs armed with a conventional MMAE warhead destroyed CD276-positive cancer cells, but were ineffective against tumor vasculature. In contrast, pyrrolobenzodiazepine-conjugated CD276-ADCs killed both cancer cells and tumor vasculature, eradicating large established tumors and metastases, and improving long-term overall survival. CD276 targeted dual-compartment ablation could aid in development of highly selective broad-acting anti-cancer therapies.

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Section: Star Methodsmentioning

confidence: 99%

Eradication of Tumors through Simultaneous Ablation of CD276/B7-H3-Positive Tumor Cells and Tumor Vasculature

Seaman

Zhu²,

Saha³

et al. 2017

Cancer Cell

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309

336

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“…These methods are based on modifying natural or engineered amino acid or carbohydrate residues in the antibody component by chemical or enzymatic reactions. Chemical site-specific conjugation of drug molecules requires the introduction of a unique chemical reactivity in the antibody, which is displayed by engineered natural, generally Cys (Casi et al, 2012; Dornan et al, 2009; Jeffrey et al, 2013; Junutula et al, 2010; Junutula et al, 2008), or engineered unnatural amino acids (Axup et al, 2012; Jackson et al, 2014; Tian et al, 2014; Zimmerman et al, 2014) or, alternatively, by engineered carbohydrates (Okeley et al, 2013; Zhou et al, 2014; Zhu et al, 2014). In certain cases, the unique chemical reactivity is generated enzymatically (Drake et al, 2014; Rabuka et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Stable and Potent Selenomab-Drug Conjugates

Nelson

Nair

et al. 2017

Cell Chemical Biology

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Summary Selenomabs are engineered monoclonal antibodies with one or more translationally incorporated selenocysteine residues. The unique reactivity of the selenol group of selenocysteine permits site-specific conjugation of drugs. Compared to other natural and unnatural amino acid and carbohydrate residues that have been used for the generation of site-specific antibody-drug conjugates, selenocysteine is particularly reactive, permitting fast, single-step, and efficient reactions under near physiological conditions. Using a tailored conjugation chemistry, we generated highly stable selenomab-drug conjugates and demonstrated their potency and selectivity in vitro and in vivo. These site-specific antibody-drug conjugates built on a selenocysteine interface revealed broad therapeutic utility in liquid and solid malignancy models.

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“…32 ELISA ELISA was performed as described previously. 16 Briefly, antigens were coated on 96-well plates at a concentration of 2 mg mL…”

Section: High-resolution Mass Spectrometrymentioning

confidence: 99%

Improving the CH1-CK heterodimerization and pharmacokinetics of 4Dm2m, a novel potent CD4-antibody fusion protein against HIV-1

Chen

Bardhi

Yang

et al. 2016

mAbs

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(2016) Improving the CH1-CK heterodimerization and pharmacokinetics of 4Dm2m, a novel potent CD4-antibody fusion protein against HIV-1, mAbs, 8:4, 761-774, DOI: 10.1080/19420862.2016 To link to this article: https://doi.org/10. 1080/19420862.2016 ABSTRACTWe previously described 4Dm2m, an exceptionally potent broadly neutralizing CD4-antibody fusion protein against HIV-1. It was generated by fusing the engineered single human CD4 domain mD1.22 to both the N and C termini of the human IgG1 heavy chain constant region and the engineered single human antibody domain m36.4, which targets the CD4-induced coreceptor binding site of the viral envelope glycoprotein, to the N terminus of the human antibody kappa light chain constant region via the (G4S)3 polypeptide linkers. However, therapeutic use of 4Dm2m was limited by its short in vivo half-life. Here, we show that a combination of three approaches have successfully increased the persistence of 4Dm2m in mice. First, to stabilize the scaffold, we enhanced heterodimerization between the heavy chain constant domain 1 (CH1) and kappa light chain constant domain (CK) by using structure-guided design and phage-display library technologies. Second, to address the possibility that long polypeptide linkers might render fusion proteins more susceptible to proteolysis, we shortened the (G4S)3 linkers or replaced them with the human IgG1 hinge sequence, which is naturally designed for both flexibility and stability. Third, we introduced two amino acid mutations into the crystallizable fragment (Fc) of the scaffold previously shown to increase antibody binding to the neonatal Fc receptor (FcRn) and prolong half-lives in vivo. Collectively, these approaches markedly increased the serum concentrations of 4Dm2m in mice while not affecting other properties of the fusion protein. The new 4Dm2m variants are promising candidates for clinical development to prevent or treat HIV-1 infection. To our knowledge, this is the first report on stabilized CH1-CK, which is potentially useful as a new heterodimerization scaffold for generation of bispecific and multispecific antibodies or proteins with a more favorable pharmacokinetic profile.

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Site-specific antibody-drug conjugation through an engineered glycotransferase and a chemically reactive sugar

Cited by 90 publications

References 37 publications

Eradication of Tumors through Simultaneous Ablation of CD276/B7-H3-Positive Tumor Cells and Tumor Vasculature

Eradication of Tumors through Simultaneous Ablation of CD276/B7-H3-Positive Tumor Cells and Tumor Vasculature

Stable and Potent Selenomab-Drug Conjugates

Improving the CH1-CK heterodimerization and pharmacokinetics of 4Dm2m, a novel potent CD4-antibody fusion protein against HIV-1

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