Development 68Ga trastuzumab Fab and bioevaluation by PET imaging in HER2/neu expressing breast cancer patients
DIR@IMTECH: CSIR-Institute of Microbial Technology
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Title |
Development 68Ga trastuzumab Fab and bioevaluation by PET imaging in HER2/neu expressing breast cancer patients
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Creator |
Rathore , Yogesh
Shukla , Jaya Laroiya, Ishita Deep , Amar Lakhanpal , Tamanna Kumar , Rajender Singh , Harmandeep Bal, Amanjit Singh , Gurpreet Thakur , Krishan Gopal Mittal , B R |
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Subject |
QR Microbiology
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Description |
Introduction: Receptors on breast cancer cells play a crucial role in the management of patients. Trastuzumab is a widely used drug for the treatment of HER2/neu expressing tumors. ImmunoPET with trastuzumab is not feasible due to slow pharmacokinetics. Fragment of antigen-binding (Fab) radiolabeled with positron emitters can be used for immunoPET. Methods: Fab has been generated by papain digestion and conjugated with the bifunctional chelating agent NOTA. The SDS-PAGE and MALDI-TOF were used to see the integrity of Fab and conjugated Fab. In-vitro stability and target specificity for HER2/neu receptors were performed in plasma and receptor binding with bio-layer interferometry (BLI) techniques. Radiolabeling was standardized with 68GaCl3 and PET imaging was performed in seven patients showing 18F fluorodeoxyglucose (18F-FDG) uptake and correlated with HER2/neu expression by immunohistochemistry. Results: Fab production was optimized at molar ratio 23:1 of trastuzumab and papain at 37 °C with a constant stirrer at 850 rpm for 22-24 h, at pH 8. Conjugation with NOTA was standardized at molar ratio 1:25 of trastuzumab Fab and NOTA. Molecular mass of trastuzumab Fab-NOTA was found approximately 46.3 kDa (~1/3 of intact antibody). Trastuzumab Fab-NOTA showed radiolabelling efficiency of 48-70% with incubation time 15 min at 37-40 °C and pH 4.5-5.0. BLI demonstrated the affinity of trastuzumab, trastuzumab Fab and trastuzumab Fab-NOTA towards HER2/neu receptor with KD of |
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Publisher |
Wolters Kluwer
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Date |
2022-04-01
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Type |
Article
PeerReviewed |
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Relation |
https://journals.lww.com/nuclearmedicinecomm/Abstract/2022/04000/Development_68Ga_trastuzumab_Fab_and_bioevaluation.11.aspx
http://crdd.osdd.net/open/2992/ |
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Identifier |
Rathore , Yogesh and Shukla , Jaya and Laroiya, Ishita and Deep , Amar and Lakhanpal , Tamanna and Kumar , Rajender and Singh , Harmandeep and Bal, Amanjit and Singh , Gurpreet and Thakur , Krishan Gopal and Mittal , B R (2022) Development 68Ga trastuzumab Fab and bioevaluation by PET imaging in HER2/neu expressing breast cancer patients. Nuclear Medicine Communications , 43 (4). pp. 458-467.
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