Influence of Histidine-Containing Tags on the Biodistribution of ADAPT Scaffold Proteins

Sarah Lindbo, Javad Garousi, Mikael Åstrand, Hadis Honarvar, Anna Orlova, Sophia Hober, Vladimir Tolmachev

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

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 99mTc(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, 99mTc or 111In. The labeling with 99mTc(CO)3 utilized the histidine-containing tags, while 111In was attached through a maleimido derivative of DOTA conjugated to the N-terminus. For 111In-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 111In-(HE)3-ADAPT6 and H6-ADAPT6 at later time points. Interestingly, in the case of 99mTc, 99mTc(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.

Original languageEnglish
Pages (from-to)716-726
Number of pages11
JournalBioconjugate Chemistry
Volume27
Issue number3
DOIs
Publication statusPublished - 16 Mar 2016
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

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