An efficient method for the site-specific <sup>99m</sup>Tc labeling of nanobody

Qi Luo; Hannan Gao; Jiyun Shi; Fan Wang

doi:10.52601/bpr.2021.210012

Volume 7 Issue 4

Sep. 2021

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Qi Luo, Hannan Gao, Jiyun Shi, Fan Wang. An efficient method for the site-specific 99mTc labeling of nanobody[J]. Biophysics Reports, 2021, 7(4): 295-303. doi: 10.52601/bpr.2021.210012

Citation:

Qi Luo, Hannan Gao, Jiyun Shi, Fan Wang. An efficient method for the site-specific ^99mTc labeling of nanobody[J]. Biophysics Reports, 2021, 7(4): 295-303. doi: 10.52601/bpr.2021.210012

Qi Luo, Hannan Gao, Jiyun Shi, Fan Wang. An efficient method for the site-specific 99mTc labeling of nanobody[J]. Biophysics Reports, 2021, 7(4): 295-303. doi: 10.52601/bpr.2021.210012

Citation:

Qi Luo, Hannan Gao, Jiyun Shi, Fan Wang. An efficient method for the site-specific ^99mTc labeling of nanobody[J]. Biophysics Reports, 2021, 7(4): 295-303. doi: 10.52601/bpr.2021.210012

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An efficient method for the site-specific ^99mTc labeling of nanobody

doi: 10.52601/bpr.2021.210012

Qi Luo¹,
Hannan Gao²,
Jiyun Shi³,
Fan Wang^{1, 2, 3, 4
,
,}

1.
Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou 510005, China
2.
Medical Isotopes Research Center and Department of Radiation Medicine, State Key Laboratory of Natural and Biomimetic Drugs, School of Basic Medical Sciences, Peking University, Beijing 100191, China
3.
Key Laboratory of Protein and Peptide Pharmaceuticals, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
4.
Beijing Translational Center for Biopharmaceuticals, Beijing 100101, China

Funds: This work was supported by National Natural Science Foundation of China (81630045, 81927802 and 81971676), and Emergency Key Program of Guangzhou Laboratory (EKPG21-16).

More Information

Corresponding author: wangfan@bjmu.edu.cn (F. Wang)
Received Date: 06 May 2021
Accepted Date: 30 June 2021
Publish Date: 17 September 2021

Abstract

Abstract

Recently, there has been a lot of interest by using nanobodies (heavy chain-only antibodies produced naturally from the Camelidae) as targeting molecules for molecular imaging, especially for the nuclear medicine imaging. A radiolabeled method that generates a homogeneous product is of utmost importance in radiotracer development for the nuclear medicine imaging. The conventional method for the radiolabeling of nanobodies is non-specifically, which conjugates the radioisotope chelating group to the side chain ɛ-amine group of lysine or sulfhydryl of cysteine of nanobodies, with a shortcoming of produce of the heterogeneous radiotracer. Here we describe a method for the site-specific radioisotope ^99mTc labeling of nanobodies by transpeptidase Sortase A. The radiolabeling process includes two steps: first step, NH₂-GGGGK(HYNIC)-COOH peptide (GGGGK = NH₂-Gly-Gly-Gly-Gly-Lys-COOH, HYNIC = 6-hydrazinonicotinyl) was labeled with ^99mTc to obtain GGGGK-HYNIC-^99mTc; second step, Sortase A catalyzes the formation of a new peptide bond between the peptide motif LPETG (NH₂-Leu-Pro-Glu-Thr-Gly-COOH) expressed C-terminally on the nanobody and the N-terminal of GGGGK-HYNIC-^99mTc. After a simple purification process, homogeneous single-conjugated and stable ^99mTc-labeled nanobodies were obtained in >50% yield. This approach demonstrates that the Sortase A-mediated conjugation is a valuable strategy for the development of site-specifically ^99mTc-labeled nanobodies.
- Nanobody,
- Sortase A,
- Site-specific radiolabeling,
- ^99mTc

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References(22)

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