Regiospecific radiolabelling of Nanofitin on Ni Magnetic Beads with [18F]FBEM and in vivo PET studies

[en] Introduction: Nanofitins are low molecular weight, single chain and cysteine-free protein scaffolds able to selectively bind a defined biological target. They derive from Sac7d bacterial protein family and are highly stable over a wide range of pH (0-13) and temperature (Tm ~80°C). Their extreme stability, low cost of production and high tolerability for chemical coupling make Nanofitins a very interesting alternative to antibodies and their fragments. Here, a hexahistidine tagged model Nanofitin (H4) directed against hen egg white lysozyme was radiolabelled and injected in mice to provide a baseline biodistribution and pharmacokinetic profiles to support future Nanofitin development programs. Method: A single cysteine residue has been genetically inserted in a model Nanofitin and its regioselective radiolabelling has been performed with 4-[18F]fluorobenzamido-N-ethylamino-maleimide ([18F]FBEM). The synthesis of [18F]FBEM has been completely implemented on a radiosynthesis unit (FastLab) including HPLC purification and formulation. Coupling with the [18F]FBEM has been achieved on a solid support (Ni magnetic beads) allowing rapid purification at room temperature without organic solvent. PET-MRI studies on C57BL/6 mice were conducted after injection of [18F]FBEM-Cys-H4 in order to access the biodistribution of this Nanofitin model. Results: Radiochemical yield (decay corrected) of 54±7% (n=4) was obtained after optimization for coupling the [18F]FBEM to Nanofitin. Pharmacokinetics results of [18F]FBEM-Cys-H4 revealed a fast clearance through the liver and the kidneys. Conclusion: An efficient new method on Ni magnetic beads was developed to radiolabelled his-tagged biomolecules with [18F]FBEM. This procedure was applied on a Nanofitin model Cys-H4 and biodistribution kinetic studies were achieved to evaluate the potential use of Nanofitin for diagnostic imaging. Fast clearance indicates that Nanofitins represent very interesting tools for diagnostic imaging.

Research center :

GIGA CRC (Cyclotron Research Center) In vivo Imaging-Aging & Memory - ULiège

Disciplines :

Chemistry

Author, co-author :

Dammicco, Sylvestre ; Université de Liège > Département de chimie (sciences) > Laboratoire de chimie organique de synthèse

Goux, Marine

Lemaire, Christian ; Université de Liège > Centre de recherches du cyclotron

Becker, Guillaume ; Université de Liège > Centre de recherches du cyclotron

Bahri, Mohamed Ali ; Université de Liège > Centre de recherche du Cyclotron

Plenevaux, Alain ; Université de Liège > Département de chimie (sciences) > Département de chimie (sciences)

Cinier, Mathieu

Luxen, André ; Université de Liège > Département de chimie (sciences) > Laboratoire de chimie organique de synthèse

Language :

English

Title :

Regiospecific radiolabelling of Nanofitin on Ni Magnetic Beads with [18F]FBEM and in vivo PET studies

Publication date :

August 2017

Journal title :

Nuclear Medicine and Biology

ISSN :

0969-8051

Publisher :

Pergamon Press, Oxford, United Kingdom

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 19 June 2017

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