Effect of Conjugation Site and Technique on the Stability and Pharmacokinetics of Antibody-Drug Conjugates

Kaempffe, Anna; Dickgießer, Stephan; Rasche, Nicolas; Bertotti, Elisa; Salve, Ilse De; Sirtori, Federico Riccardi; Kellner, Roland; Könning, Doreen; Hecht, Stefan; Anderl, Jan; Kolmar, Harald; Schröter, Christian

doi:10.1016/j.xphs.2021.08.002

Cited by 17 publications

(11 citation statements)

References 57 publications

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“…Plasma concentrations of the molecules were measured via ELISA using a ligand-binding assay with a lower limit of quantification of 50 ng/mL as described previously. 51 Biotin SP-conjugated AffiniPure goat anti-human IgG (Fcγ fragment specific, Jackson Immuno Research, 109-065-098) was used as capture reagent and SULFO-tag labeled AffiniPure goat anti-human IgG (Fcγ fragment specific, Jackson Immuno Research, 109-005-098) was used as a detection reagent.…”

Section: Methodsmentioning

confidence: 99%

Impact of antibody architecture and paratope valency on effector functions of bispecific NKp30 x EGFR natural killer cell engagers

Boje,

Pekar,

Koep

et al. 2024

mAbs

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Natural killer (NK) cells emerged as a promising effector population that can be harnessed for anti-tumor therapy. In this work, we constructed NK cell engagers (NKCEs) based on NKp30-targeting single domain antibodies (sdAbs) that redirect the cytotoxic potential of NK cells toward epidermal growth factor receptor (EGFR)-expressing tumor cells. We investigated the impact of crucial parameters such as sdAb location, binding valencies, the targeted epitope on NKp30, and the overall antibody architecture on the redirection capacity. Our study exploited two NKp30-specific sdAbs, one of which binds a similar epitope on NKp30 as its natural ligand B7-H6, while the other sdAb addresses a non-competing epitope. For EGFR-positive tumor targeting, humanized antigen-binding domains of therapeutic antibody cetuximab were used. We demonstrate that NKCEs bivalently targeting EGFR and bivalently engaging NKp30 are superior to monovalent NKCEs in promoting NK cell-mediated tumor cell lysis and that the architecture of the NKCE can substantially influence killing capacities depending on the NKp30-targeting sdAb utilized. While having a pronounced impact on NK cell killing efficacy, the capabilities of triggering antibody-dependent cellular phagocytosis or complement-dependent cytotoxicity were not significantly affected comparing the bivalent IgG-like NKCEs with cetuximab. However, the fusion of sdAbs can have a slight impact on the NK cell release of immunomodulatory cytokines, as well as on the pharmacokinetic profile of the NKCE due to unfavorable spatial orientation within the molecule architecture. Ultimately, our findings reveal novel insights for the engineering of potent NKCEs triggering the NKp30 axis.

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Section: Methodsmentioning

confidence: 99%

Impact of antibody architecture and paratope valency on effector functions of bispecific NKp30 x EGFR natural killer cell engagers

Boje,

Pekar,

Koep

et al. 2024

mAbs

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“…Serum Stability. The serum stability assay was conducted as previously described 29,35,36 Cancer Cell Cytotoxicity Assay. Human cancer cell lines were obtained from the American Type Culture Collection (CEA5-positive: SK-CO-1, MKN-45, LS174T; CEA5-negative MDA-MB-231) and maintained according to standard culture conditions (SK-CO-1, MKN-45: 37 °C, 5% CO 2 , 95% humidity; LS174T, MDA-MB-231: 37 °C, 10% CO 2 , 95% humidity).…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

“…The serum stability assay was conducted as previously described ,, and upon applying some minor modifications. Exatecan antibody-drug conjugates were incubated at a final concentration of 5 μM conjugated exatecan (considering the DAR of each construct) in human, mouse, and cyno serum.…”

Section: Methodsmentioning

confidence: 99%

Generation and Characterization of Iduronidase-Cleavable ADCs

Jäger,

Könning,

Rasche

et al. 2023

Bioconjugate Chem.

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“…This stochastic distribution leads to unpredictable pharmacokinetic effects, as for example, binding of the payload to sites on the antibody that participate in antigen binding can substantially alter the pharmacokinetics and biological activity of the ADC [ 28 ]. In vitro assays and pharmacokinetic analyses in xenograft models have introduced cysteine conjugation at various antibody positions and compared this to enzymatic conjugation using microbial transglutaminase on the light chain or heavy chain [ 29 ]. ADCs produced using enzymatic conjugation to the light chain or position Q295 on the antibody had superior pharmacokinetic behaviour, as did those engineered with cystine conjugation to the L328 position [ 29 ].…”

Section: Adc Structure and Mechanism Of Actionmentioning

confidence: 99%

Antibody–Drug Conjugates: Ushering in a New Era of Cancer Therapy

2023

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Antibody–drug conjugates (ADCs) have provided new therapeutic options and significant promise for patients with cancer, particularly where existing treatments are limited. Substantial effort in ADC development is underway globally, with 13 ADCs currently approved and many more in development. The therapeutic benefits of ADCs leverage the ability to selectively target cancer cells through antibody binding, resultant relative sparing of non-malignant tissues, and the targeted delivery of a cytotoxic payload. Consequently, this drug class has demonstrated activity in multiple malignancies refractory to standard therapeutic options. Despite this, limitations exist, including narrow therapeutic windows, unique toxicity profiles, development of therapeutic resistance, and appropriate biomarker selection. This review will describe the development of ADCs, their mechanisms of action, pivotal trials, and approved indications and identify common themes. Current challenges and opportunities will be discussed for this drug class in cancer therapeutics at a time when significant developments in antibody therapies, immunotherapy, and targeted agents are occurring.

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Effect of Conjugation Site and Technique on the Stability and Pharmacokinetics of Antibody-Drug Conjugates

Cited by 17 publications

References 57 publications

Impact of antibody architecture and paratope valency on effector functions of bispecific NKp30 x EGFR natural killer cell engagers

Impact of antibody architecture and paratope valency on effector functions of bispecific NKp30 x EGFR natural killer cell engagers

Generation and Characterization of Iduronidase-Cleavable ADCs

Antibody–Drug Conjugates: Ushering in a New Era of Cancer Therapy

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