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Optimizing and determining the click chemistry mediated Cu-64 radiolabeling and physiochemical characteristics of trastuzumab conjugates.
Bhise, Abhinav; Park, Hyun; Rajkumar, Subramani; Lee, Kiwoong; Cho, Seong Hwan; Lim, Jeong Eun; Kim, Jung Young; Lee, Kyo Chul; Yoon, Young-Ran; Yoo, Jeongsoo.
Afiliación
  • Bhise A; Department of Molecular Medicine, Brain Korea 21, Four KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, School of Medicine, Kyungpook National University, Daegu, 41944, Republic of Korea.
  • Park H; Division of Applied RI, Korea Institute of Radiological and Medical Sciences, Seoul, 01812, Republic of Korea.
  • Rajkumar S; Department of Molecular Medicine, Brain Korea 21, Four KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, School of Medicine, Kyungpook National University, Daegu, 41944, Republic of Korea.
  • Lee K; Department of Molecular Medicine, Brain Korea 21, Four KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, School of Medicine, Kyungpook National University, Daegu, 41944, Republic of Korea.
  • Cho SH; Department of Molecular Medicine, Brain Korea 21, Four KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, School of Medicine, Kyungpook National University, Daegu, 41944, Republic of Korea.
  • Lim JE; Department of Molecular Medicine, Brain Korea 21, Four KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, School of Medicine, Kyungpook National University, Daegu, 41944, Republic of Korea.
  • Kim JY; Division of Applied RI, Korea Institute of Radiological and Medical Sciences, Seoul, 01812, Republic of Korea.
  • Lee KC; Division of Applied RI, Korea Institute of Radiological and Medical Sciences, Seoul, 01812, Republic of Korea.
  • Yoon YR; Department of Molecular Medicine, Brain Korea 21, Four KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, School of Medicine, Kyungpook National University, Daegu, 41944, Republic of Korea.
  • Yoo J; Department of Molecular Medicine, Brain Korea 21, Four KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, School of Medicine, Kyungpook National University, Daegu, 41944, Republic of Korea. Electronic address: yooj@knu.ac.kr.
Biochem Biophys Res Commun ; 638: 28-35, 2023 01 01.
Article en En | MEDLINE | ID: mdl-36436339
Over the last decade, 64Cu-labeling of monoclonal antibody (mAb) via inverse electron demand Diels-Alder click chemistry (IEDDA) have received much attention. Despite the tetrazine-transcyclooctene (Tz-TCO) click chemistry's convenience and efficiency in mAb labeling, there is limited information about the ideal parameters in the development of click chemistry mediated (radio)immunoconjugates. This encourages us to conduct a systematic optimization while concurrently determining the physiochemical characteristics of the model mAb, trastuzumab, and TCO conjugates. To accomplish this, we investigated a few critical parameters, first, we determined the degree of conjugations with varying molar equivalents (eq.) of TCO (3, 5, 10, and 15 eq.). Through analytical techniques like size exclusion chromatography, dynamic light scattering, and zeta potential, qualitative analysis were performed to determine the purity, degree of aggregation and net charge of the conjugates. We found that as the degree of conjugation increased the purity of intact mAb fraction is compromised and net charge of conjugates became less positive. Next, all trastuzumab-PEG4-TCO conjugates with varying molar ratio and quantity (30, 50, 100, 200, 250 µg) were radiolabeled with 64Cu-NOTA-PEG4-Tz via IEDDA click chemistry and radiochemical yields were determined by radio-thin layer chromatography. The radiochemical yields of trastuzumab conjugates improved with increased amount and molar ratio. Next, we investigated the effect of the radioprotectant ascorbic acid (AA) of varied concentrations (0.25, 0.5, 0.75, 1 mM) on radiochemical yields and subsequent pharmacokinetics. A concentration of 0.25 mM of AA was found to be optimal for click reaction and in vivo biodistribution. Finally, we investigated the indirect influence of bioconjugation buffers on radiochemical yields and biodistribution in NIH3T6.7 tumor models that resulted approximately ∼11 %ID/g tumor uptake.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Radioisótopos de Cobre / Neoplasias Tipo de estudio: Prognostic_studies / Qualitative_research Límite: Humans Idioma: En Revista: Biochem Biophys Res Commun Año: 2023 Tipo del documento: Article Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Radioisótopos de Cobre / Neoplasias Tipo de estudio: Prognostic_studies / Qualitative_research Límite: Humans Idioma: En Revista: Biochem Biophys Res Commun Año: 2023 Tipo del documento: Article Pais de publicación: Estados Unidos