Radiolabeling of porcine, murine growth hormone and a potential antagonist G118R-mGH for biodistribution study

Amanda Palermo Nunes; Filipe Menezes; Larissa Andrade Almeida; Stephanie A.  Pomin; Patricia Lima Falcão; Carlos Soares

doi:10.15392/2319-0612.2024.2744

Autores/as

Amanda Palermo Nunes IPEN https://orcid.org/0009-0004-9684-9507 (no autenticado)
Filipe Menezes IPEN https://orcid.org/0009-0001-7830-0447 (no autenticado)
Larissa Andrade Almeida IPEN https://orcid.org/0000-0002-0354-8384 (no autenticado)
Stephanie A. Pomin IPEN https://orcid.org/0000-0001-5637-2303 (no autenticado)
Patricia Lima Falcão IPEN https://orcid.org/0000-0003-4640-9939 (no autenticado)
Carlos Soares IPEN https://orcid.org/0000-0002-7982-1789 (no autenticado)

DOI:

https://doi.org/10.15392/2319-0612.2024.2744

Palabras clave:

Growth Hormone , Radiolabeling, Iodine, SPECT-CT

Resumen

This study aimed to radiolabel porcine growth hormone (pGH), murine growth hormone (mGH), and its antagonist (G118R-mGH) with the iodine radioisotopes ¹³¹I and ¹²³I, to investigate the biodistribution and brain transport of these hormones. Radiolabeling was performed using the Chloramine T method, a protein iodination technique. The radiolabeled products were characterized by physicochemical techniques such as SDS-PAGE, size-exclusion high-performance liquid chromatography (HPLC-SE), and ascending paper chromatography, with measurements performed using single-photon emission computed tomography (SPECT). These measurements demonstrated high radiochemical purity, exceeding 95%, and preservation of molecular size. In vitro stability assays indicated that the radiotracers maintained their integrity for at least 24 hours. In vivo biodistribution studies in mice revealed distinct tissue distribution patterns for mGH and G118R-mGH, suggesting different uptake and metabolism mechanisms. The use of ¹²³I allowed for SPECT-CT imaging studies, which proved important for assessing the ability of mGH and G118R-mGH to cross the blood-brain barrier and distribute in specific regions of the brain. The results obtained in this work highlight the versatility of the radiolabeling protocol employed and its potential for investigating complex biological processes and application in research.

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Biografía del autor/a

Carlos Soares, IPEN

Graduado em Farmácia e Bioquímica pela Universidade de São Paulo (1989), realizou Mestrado (1995) e Doutorado (2000) em Tecnologia Nuclear - Aplicações pela Universidade de São Paulo. Atualmente é pesquisador do Instituto de Pesquisas Energéticas e Nucleares (IPEN-CNEN/SP) e professor de pós-graduação vinculado à Universidade de São Paulo. Com experiência em biotecnologia na expressão de proteínas recombinantes por bactéria e por células de mamífero. Atua principalmente no seguinte tema: síntese, purificação, caracterização e aplicações de hormônios recombinantes. Atualmente é Gerente do Centro de Biotecnologia do IPEN. Bolsista de Produtividade desen. Tec. e Extensão Inovadora do CNPq - Nível 2.

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