Discovery, optimization and biodistribution of an Affibody molecule for imaging of CD69

Abstract Due to the wide scale of inflammatory processes in different types of disease, more sensitive and specific biomarkers are required to improve prevention and treatment. Cluster of differentiation 69 (CD69) is one of the earliest cell surface proteins expressed by activated leukocytes. Here w...

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Autores principales:	Jonas Persson, Emmi Puuvuori, Bo Zhang, Irina Velikyan, Ola Åberg, Malin Müller, Per-Åke Nygren, Stefan Ståhl, Olle Korsgren, Olof Eriksson, John Löfblom
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b6d61dc149314cc4990935ca6c6c382f
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spelling	oai:doaj.org-article:b6d61dc149314cc4990935ca6c6c382f2021-12-02T17:37:40ZDiscovery, optimization and biodistribution of an Affibody molecule for imaging of CD6910.1038/s41598-021-97694-62045-2322https://doaj.org/article/b6d61dc149314cc4990935ca6c6c382f2021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-97694-6https://doaj.org/toc/2045-2322Abstract Due to the wide scale of inflammatory processes in different types of disease, more sensitive and specific biomarkers are required to improve prevention and treatment. Cluster of differentiation 69 (CD69) is one of the earliest cell surface proteins expressed by activated leukocytes. Here we characterize and optimize potential new imaging probes, Affibody molecules targeting CD69 for imaging of activated immune cells. Analysis of candidates isolated in a previously performed selection from a Z variant E. coli library to the recombinant extracellular domain of human CD69, identified one cross-reactive Z variant with affinity to murine and human CD69. Affinity maturation was performed by randomization of the primary Z variant, followed by selections from the library. The resulting Z variants were evaluated for affinity towards human and murine CD69 and thermal stability. The in vivo biodistribution was assessed by SPECT/CT in rats following conjugation of the Z variants by a DOTA chelator and radiolabeling with Indium-111. A primary Z variant with a Kd of approximately 50 nM affinity to human and murine CD69 was identified. Affinity maturation generated 5 additional Z variants with improved or similar affinity. All clones exhibited suitable stability. Radiolabeling and in vivo biodistribution in rat demonstrated rapid renal clearance for all variants, while the background uptake and washout varied. The variant ZCD69:4 had the highest affinity for human and murine CD69 (34 nM) as well as the lowest in vivo background binding. In summary, we describe the discovery, optimization and evaluation of novel Affibody molecules with affinity for CD69. Affibody molecule ZCD69:4 is suitable for further development for imaging of activated immune cells.Jonas PerssonEmmi PuuvuoriBo ZhangIrina VelikyanOla ÅbergMalin MüllerPer-Åke NygrenStefan StåhlOlle KorsgrenOlof ErikssonJohn LöfblomNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jonas Persson Emmi Puuvuori Bo Zhang Irina Velikyan Ola Åberg Malin Müller Per-Åke Nygren Stefan Ståhl Olle Korsgren Olof Eriksson John Löfblom Discovery, optimization and biodistribution of an Affibody molecule for imaging of CD69
description	Abstract Due to the wide scale of inflammatory processes in different types of disease, more sensitive and specific biomarkers are required to improve prevention and treatment. Cluster of differentiation 69 (CD69) is one of the earliest cell surface proteins expressed by activated leukocytes. Here we characterize and optimize potential new imaging probes, Affibody molecules targeting CD69 for imaging of activated immune cells. Analysis of candidates isolated in a previously performed selection from a Z variant E. coli library to the recombinant extracellular domain of human CD69, identified one cross-reactive Z variant with affinity to murine and human CD69. Affinity maturation was performed by randomization of the primary Z variant, followed by selections from the library. The resulting Z variants were evaluated for affinity towards human and murine CD69 and thermal stability. The in vivo biodistribution was assessed by SPECT/CT in rats following conjugation of the Z variants by a DOTA chelator and radiolabeling with Indium-111. A primary Z variant with a Kd of approximately 50 nM affinity to human and murine CD69 was identified. Affinity maturation generated 5 additional Z variants with improved or similar affinity. All clones exhibited suitable stability. Radiolabeling and in vivo biodistribution in rat demonstrated rapid renal clearance for all variants, while the background uptake and washout varied. The variant ZCD69:4 had the highest affinity for human and murine CD69 (34 nM) as well as the lowest in vivo background binding. In summary, we describe the discovery, optimization and evaluation of novel Affibody molecules with affinity for CD69. Affibody molecule ZCD69:4 is suitable for further development for imaging of activated immune cells.
format	article
author	Jonas Persson Emmi Puuvuori Bo Zhang Irina Velikyan Ola Åberg Malin Müller Per-Åke Nygren Stefan Ståhl Olle Korsgren Olof Eriksson John Löfblom
author_facet	Jonas Persson Emmi Puuvuori Bo Zhang Irina Velikyan Ola Åberg Malin Müller Per-Åke Nygren Stefan Ståhl Olle Korsgren Olof Eriksson John Löfblom
author_sort	Jonas Persson
title	Discovery, optimization and biodistribution of an Affibody molecule for imaging of CD69
title_short	Discovery, optimization and biodistribution of an Affibody molecule for imaging of CD69
title_full	Discovery, optimization and biodistribution of an Affibody molecule for imaging of CD69
title_fullStr	Discovery, optimization and biodistribution of an Affibody molecule for imaging of CD69
title_full_unstemmed	Discovery, optimization and biodistribution of an Affibody molecule for imaging of CD69
title_sort	discovery, optimization and biodistribution of an affibody molecule for imaging of cd69
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/b6d61dc149314cc4990935ca6c6c382f
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Discovery, optimization and biodistribution of an Affibody molecule for imaging of CD69

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