Selection of an optimal cysteine-containing peptide-based chelator for labeling of affibody molecules with 188Re

Altai, Mohamed; Honarvar, Hadis; Wållberg, Helena; Strand, Joanna; Varasteh, Zohreh; Rosestedt, Maria; Orlova, Anna; Dunås, Finn; Sandström, Mattias; Löfblom, John; Tolmachev, Vladimir; Ståhl, Stefan

doi:10.1016/j.ejmech.2014.09.082

Cited by 20 publications

(14 citation statements)

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“…This indicated that the main reason of the release was re-oxidation of technetium. This was not what we have observed earlier for anti-HER2 and anti-IGF-1R affibody molecules, where even -GGGC provided a stable attachment of 99m Tc and 188 Re (Wållberg et al 2011; Altai et al 2012, 2014a, b; Mitran et al 2015). A possible explanation for the current results is that some side chains in the affibody molecules form an alternative chelating pocket and a part of technetium is bound not only to a cysteine-containing chelator, but also to this much weaker site.…”

Section: Discussioncontrasting

confidence: 67%

Influence of composition of cysteine-containing peptide-based chelators on biodistribution of 99mTc-labeled anti-EGFR affibody molecules

et al. 2018

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Epidermal growth factor receptor (EGFR) is overexpressed in a number of cancers and is the molecular target for several anti-cancer therapeutics. Radionuclide molecular imaging of EGFR expression should enable personalization of anti-cancer treatment. Affibody molecule is a promising type of high-affinity imaging probes based on a non-immunoglobulin scaffold. A series of derivatives of the anti-EGFR affibody molecule ZEGFR:2377, having peptide-based cysteine-containing chelators for conjugation of 99mTc, was designed and evaluated. It was found that glutamate-containing chelators Gly-Gly-Glu-Cys (GGEC), Gly-Glu-Glu-Cys (GEEC) and Glu-Glu-Glu-Cys (EEEC) provide the best labeling stability. The glutamate containing conjugates bound to EGFR-expressing cells specifically and with high affinity. Specific targeting of EGFR-expressing xenografts in mice was demonstrated. The number of glutamate residues in the chelator had strong influence on biodistribution of radiolabeled affibody molecules. Increase of glutamate content was associated with lower uptake in normal tissues. The 99mTc-labeled variant containing the EEEC chelator provided the highest tumor-to-organ ratios. In conclusion, optimizing the composition of peptide-based chelators enhances contrast of imaging of EGFR-expression using affibody molecules.Electronic supplementary materialThe online version of this article (10.1007/s00726-018-2571-1) contains supplementary material, which is available to authorized users.

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Section: Discussioncontrasting

confidence: 67%

Influence of composition of cysteine-containing peptide-based chelators on biodistribution of 99mTc-labeled anti-EGFR affibody molecules

et al. 2018

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“…The present study evaluated preconditions to labeling of affibody molecule-derived imaging probes using peptide-based cysteine-containing chelators. Chelators of that type provided a stable labeling of anti-HER2 affibody molecules with 99m Tc and 188 Re (42,52). An apparent advantage of such chelators is that they are ‘built in’ in the recombinantly produced affibody molecule, and there is no need in additional steps of coupling of the chelator and purification of the conjugate.…”

Section: Discussionmentioning

confidence: 99%

“…An apparent advantage of such chelators is that they are ‘built in’ in the recombinantly produced affibody molecule, and there is no need in additional steps of coupling of the chelator and purification of the conjugate. In addition, selection of amino acids in the chelator may enable a fine-tuning of residualizing properties of the label (52–54). However, there was a risk of losing site-specificity of the label when amino acid composition of an affibody molecules was modified (43).…”

Section: Discussionmentioning

confidence: 99%

Feasibility of imaging of epidermal growth factor receptor expression with ZEGFR:2377 affibody molecule labeled with 99mTc using a peptide-based cysteine-containing chelator

Andersson

Oroujeni

Garousi

et al. 2016

International Journal of Oncology

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The epidermal growth factor receptor (EGFR) is overexpressed in a number of malignant tumors and is a molecular target for several specific anticancer antibodies and tyrosine kinase inhibitors. The overexpression of EGFR is a predictive biomarker for response to several therapy regimens. Radionuclide molecular imaging might enable detection of EGFR overexpression by a non-invasive procedure and could be used repeatedly. Affibody molecules are engineered scaffold proteins, which could be selected to have a high affinity and selectivity to predetermined targets. The anti-EGFR ZEGFR:2377 affibody molecule is a potential imaging probe for EGFR detection. The use of the generator-produced radionuclide 99mTc should facilitate clinical translation of an imaging probe due to its low price, availability and favorable dosimetry of the radionuclide. In the present study, we evaluated feasibility of ZEGFR:2377 labeling with 99mTc using a peptide-based cysteine-containing chelator expressed at the C-terminus of ZEGFR:2377. The label was stable in vitro under cysteine challenge. In addition, 99mTc-ZEGFR:2377 was capable of specific binding to EGFR-expressing cells with high affinity (274 pM). Studies in BALB/C nu/nu mice bearing A431 xenografts demonstrated that 99mTc-ZEGFR:2377 accumulates in tumors in an EGFR-specific manner. The tumor uptake values were 3.6±1 and 2.5±0.4% ID/g at 3 and 24 h after injection, respectively. The corresponding tumor-to-blood ratios were 1.8±0.4 and 8±3. The xenografts were clearly visualized at both time-points. This study demonstrated the potential of 99mTc-labeled ZEGFR:2377 for imaging of EGFR in vivo.

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“…In order to deliver the required dose to tumors without exceeding the limiting dose to the kidneys (23 Gy), several strategies have been developed. The use of labeling approaches that provide nonresidualizing radiocatabolites, such as radioiodination [5,6] or the use of some peptide-based chelators for rhenium radioisotopes and technetium-99m [12][13][14][15][16][17], was shown to decrease the retention of activity in the kidneys by several folds and to improve tumor-to-kidney ratios. Another effective strategy is the fusion of an ESP to an albumin-binding domain (ABD).…”

Section: Introductionmentioning

confidence: 99%

On the prevention of kidney uptake of radiolabeled DARPins

et al. 2020

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Background: Designed ankyrin repeat proteins (DARPins) are small engineered scaffold proteins (14-18 kDa) that demonstrated promising tumor-targeting properties in preclinical studies. However, high renal accumulation of activity for DARPins labeled with residualizing labels is a limitation for targeted radionuclide therapy. A better understanding of the mechanisms behind the kidney uptake of DARPins could aid the development of strategies to reduce it. In this study, we have investigated whether the renal uptake of [ 99m Tc]Tc(CO) 3-G3 DARPin could be reduced by administration of compounds that act on various parts of the reabsorption system in the kidney. Results: Co-injection of lysine or Gelofusine was not effective for the reduction of kidney uptake of [ 99m Tc]Tc(CO) 3-G3. Administration of sodium maleate before the injection of [ 99m Tc]Tc(CO) 3-G3 reduced the kidney-associated activity by 60.4 ± 10.3%, while administration of fructose reduced it by 46.9 ± 7.6% compared with the control. The decrease in the kidney uptake provided by sodium maleate was also observed for [ 99m Tc]Tc(CO) 3-9_29 DARPin. Preinjection of colchicine, probenecid, mannitol, or furosemide had no effect on the kidney uptake of [ 99m Tc]Tc(CO) 3-G3. Kidney autoradiography showed mainly cortical accumulation of activity for all studied groups. Conclusion: Common clinical strategies were not effective for the reduction of kidney uptake of [ 99m Tc]Tc(CO) 3-G3. Both fructose and maleate lower the cellular ATP level in the proximal tubule cells and their reduction of the kidney reuptake indicates the involvement of an ATP-driven uptake mechanism. The decrease provided by maleate for both G3 and 9_29 DARPins indicates that their uptake proceeds through a mechanism independent of DARPin structure and binding site composition.

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Selection of an optimal cysteine-containing peptide-based chelator for labeling of affibody molecules with 188Re

Cited by 20 publications

References 46 publications

Influence of composition of cysteine-containing peptide-based chelators on biodistribution of 99mTc-labeled anti-EGFR affibody molecules

Influence of composition of cysteine-containing peptide-based chelators on biodistribution of 99mTc-labeled anti-EGFR affibody molecules

Feasibility of imaging of epidermal growth factor receptor expression with ZEGFR:2377 affibody molecule labeled with 99mTc using a peptide-based cysteine-containing chelator

On the prevention of kidney uptake of radiolabeled DARPins

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