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Influence of Histidine-Containing Tags on the Biodistribution of ADAPT Scaffold Proteins.
KTH Royal Inst Technol, Dept Prot Technol, SE-10691 Stockholm, Sweden.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Medical Radiation Science. (Vladimir Tolmachev)
KTH Royal Inst Technol, Dept Prot Technol, SE-10691 Stockholm, Sweden.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Medical Radiation Science. (Vladimir Tolmachev)
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2016 (English)In: Bioconjugate chemistry, ISSN 1043-1802, E-ISSN 1520-4812, Vol. 27, no 3, 716-726 p.Article in journal (Refereed) Published
Abstract [en]

Engineered scaffold proteins (ESP) are high-affinity binders that can be used as probes for radionuclide imaging. Histidine-containing tags enable both efficient purification of ESP and radiolabeling with (99m)Tc(CO)3. Earlier studies demonstrated that the use of a histidine-glutamate-histidine-glutamate-histidine-glutamate (HE)3-tag instead of the commonly used hexahistidine (H6)-tag reduces hepatic uptake of radiolabeled ESP and short peptides. Here, we investigated the influence of histidine-containing tags on the biodistribution of a novel type of ESP, ADAPTs. A series of anti-HER2 ADAPT probes having H6- or (HE)3-tags in the N-termini were prepared. The constructs, (HE)3-ADAPT6 and H6-ADAPT6, were labeled with two different nuclides, (99m)Tc or (111)In. The labeling with (99m)Tc(CO)3 utilized the histidine-containing tags, while (111)In was attached through a maleimido derivative of DOTA conjugated to the N-terminus. For (111)In-labeled ADAPTs, the use of (HE)3 provided a significantly (p < 0.05) lower hepatic uptake at 1 h after injection, but there was no significant difference in hepatic uptake of (111)In-(HE)3-ADAPT6 and H6-ADAPT6 at later time points. Interestingly, in the case of (99m)Tc, (99m)Tc(CO)3-H6-ADAPT6 provided significantly (p < 0.05) lower uptake in a number of normal tissues and was more suitable as an imaging probe. Thus, the influence of histidine-containing tags on the biodistribution of the novel ADAPT scaffold proteins was different compared to its influence on other ESPs studied so far. Apparently, the effect of a histidine-containing tag on the biodistribution is highly dependent on the scaffold composition of the ESP.

Place, publisher, year, edition, pages
2016. Vol. 27, no 3, 716-726 p.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-286833DOI: 10.1021/acs.bioconjchem.5b00677ISI: 000372478600026PubMedID: 26781756OAI: oai:DiVA.org:uu-286833DiVA: diva2:922065
Funder
Swedish Cancer Society, CAN 2015/350Swedish Research Council, 2015-02353, 621-2012-5088
Note

First two authors (Lindbo and Garousi) contributed equally

Available from: 2016-04-21 Created: 2016-04-21 Last updated: 2017-11-30Bibliographically approved
In thesis
1. Development of ADAPT-based tracers for radionuclide molecular imaging of cancer
Open this publication in new window or tab >>Development of ADAPT-based tracers for radionuclide molecular imaging of cancer
2017 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

ABD-Derived Affinity Proteins (ADAPTs) is a novel class of small engineered scaffold proteins based on albumin-binding domain (ABD) of streptococcal protein G. High affinity ADAPT  binders against various therapeutic targets can be selected.  In this thesis, we report a development of ADAPT-based radionuclide imaging agents providing high sensitivity and specificity of molecular imaging of HER2 expression in disseminated cancers.

We investigated the feasibility of the use of ADAPTs as imaging agents and influence of molecular design and radiolabeling chemistry on in vivo targeting and biodistribution properties of the tracers.

In Paper I we demonstrated the feasibility of the use of anti-HER2 ADAPT6 molecule as a high contrast imaging agent;

In Paper II we evaluated the influence of composition of histidine-containing tag on in vivo biodistribution of ADAPT-based tracers labeled with 99mTc using 99mTc(CO)3 binding to histidine-containing tags and 111In using DOTA chelator at N-terminus;

In Paper III we evaluated the influence of different aspects of N-terminus leading sequence on targeting including effect of sequence size on clearance rate and effect of the composition of the sequence on biodistribution profile;

In Paper IV, we evaluated the influence of residualizing properties and positioning of the label on biodistribution and targeting; and

In Paper V, we compared tumor-targeting properties of the ADAPT6 labeled at C-terminus with 99mTc using N3S chelator and 111In using DOTA chelator.

In conclusion, ADAPTs constitute a very promising class of targeting probes for molecular imaging providing high contrast. Molecular design of the ADAPT proteins and chelators/linkers for labeling has an appreciable effect on their imaging properties.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2017. 99 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 1354
Keyword
ADAPT, scaffold protein, albumin-binding domain (ABD), Affinity protein, HER2, radionuclide molecular imaging
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-327419 (URN)978-91-513-0030-6 (ISBN)
Public defence
2017-09-29, Fåhraeus Hall, Rudbecklaboratoriet, Dag Hammarskjölds väg 20, Uppsala, 13:00 (English)
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Available from: 2017-09-06 Created: 2017-08-10 Last updated: 2017-09-08

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Garousi, JavadHonarvar, HadisOrlova, AnnaTolmachev, Vladimir

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