Leveraging a dual variable domain immunoglobulin to create a site-specifically modified radioimmunoconjugate Journal Article
| Authors: | MacPherson, D. S.; Hwang, D.; Sarrett, S. M.; Keinänen, O.; Rodriguez, C.; Rader, C.; Zeglis, B. M. |
| Article Title: | Leveraging a dual variable domain immunoglobulin to create a site-specifically modified radioimmunoconjugate |
| Abstract: | Site-specifically modified radioimmunoconjugates exhibit superior in vitro and in vivo behavior compared to analogues synthesized via traditional stochastic methods. However, the development of approaches to site-specific bioconjugation that combine high levels of selectivity, simple reaction conditions, and clinical translatability remains a challenge. Herein, we describe a novel solution to this problem: the use of dual-variable domain immunoglobulins (DVD-IgG). More specifically, we report the synthesis, in vitro evaluation, and in vivo validation of a 177Lu-labeled radioimmunoconjugate based on HER2DVD, a DVD-IgG containing the HER2-targeting variable domains of trastuzumab and the catalytic variable domains of IgG h38C2. To this end, we first modified HER2DVD with a phenyloxadiazolyl methlysulfone-modified variant of the chelator CHX-A′′-DTPA (PODS-CHX-A′′-DTPA) and verified the site-specificity of the conjugation for the reactive lysines within the catalytic domains via chemical assay, MALDI-ToF mass spectrometry, and SDS-PAGE. The chelator-bearing immunoconjugate was subsequently labeled with [177Lu]Lu3+to produce the completed radioimmunoconjugate, [177Lu]Lu-CHX-A′′-DTPAPODS-HER2DVD, in >80% radiochemical conversion and a specific activity of 29.5 ± 7.1 GBq/μmol. [177Lu]Lu-CHX-A′′-DTPAPODS-HER2DVD did not form aggregates upon prolonged incubation in human serum, displayed 87% stability to demetalation over a 7 days of incubation in serum, and exhibited an immunoreactive fraction of 0.95 with HER2-coated beads. Finally, we compared the pharmacokinetic profile of [177Lu]Lu-CHX-A′′-DTPAPODS-HER2DVD to that of a 177Lu-labeled variant of trastuzumab in mice bearing subcutaneous HER2-expressing BT-474 human breast cancer xenografts. The in vivo performance of [177Lu]Lu-CHX-A′′-DTPAPODS-HER2DVD matched that of 177Lu-labeled trastuzumab, with the former producing a tumoral activity concentration of 34.1 ± 12.1 %ID/g at 168 h and tumor-to-blood, tumor-to-liver, and tumor-to-kidney activity concentration ratios of 10.5, 9.6, and 21.8, respectively, at the same time point. Importantly, the DVD-IgG did not exhibit a substantially longer serum half-life than the traditional IgG despite its significantly larger size (202 kDa for the former vs 148 kDa for the latter). Taken together, these data suggest that DVD-IgGs represent a viable platform for the future development of highly effective site-specifically labeled radioimmunoconjugates for diagnostic imaging, theranostic imaging, and radioimmunotherapy. © 2023 American Chemical Society. All rights reserved. |
| Keywords: | mouse; animal; animals; mice; cell line, tumor; breast neoplasms; chemistry; immunoglobulin g; breast tumor; tumor cell line; trastuzumab; radioimmunotherapy; chelating agents; her2; antibody conjugate; immunoconjugates; pentetic acid; chelating agent; humans; human; female; targeted radionuclide therapy; 177lu; site-specific bioconjugation; catalytic antibody; dual-variable domain antibody; site-selective bioconjugation |
| Journal Title: | Molecular Pharmaceutics |
| Volume: | 20 |
| Issue: | 1 |
| ISSN: | 1543-8384 |
| Publisher: | American Chemical Society |
| Date Published: | 2023-01-02 |
| Start Page: | 775 |
| End Page: | 782 |
| Language: | English |
| DOI: | 10.1021/acs.molpharmaceut.2c00700 |
| PUBMED: | 36377696 |
| PROVIDER: | scopus |
| PMCID: | PMC10263003 |
| DOI/URL: | |
| Notes: | Article -- MSK Cancer Center Support Grant (P30 CA008748) acknowledged in PDF -- MSK corresponding author is Brian Zeglis -- Export Date: 1 February 2023 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
MSK Authors
Related MSK Work