Phage display selection of Affibody molecules with specific binding to the extracellular domain of the epidermal growth factor receptor

Friedman, Mikaela; Nordberg, Erika; Höidén-Guthenberg, Ingmarie; Brismar, Hjalmar; Adams, Gregory P.; Nilsson, Fred; Carlsson, Jörgen; Ståhl, Stefan

doi:10.1093/protein/gzm011

Cited by 107 publications

(107 citation statements)

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“…The dimer format of each affibody molecule in the bs affibody protein will most likely give an additional increase in functional affinity (avidity), as demonstrated previously [38,39]. Furthermore, high selectivity of the Z HER2:342 and Z EGFR:1907 affibody molecules to their respective target protein has been shown in previous studies [30,39]. In the present study, binding to HER2 and EGFR was confirmed for the bs affibody protein (Figure 2).…”

Section: Single and Dual Binding Biosensor Analysissupporting

confidence: 86%

“…The affinities (equilibrium dissociation constant, K D ) of the monomeric affibody molecules were previously determined to be 22 pM for Z HER2:342 [30] and 5.4 nM for Z EGFR:1907 [31]. The dimer format of each affibody molecule in the bs affibody protein will most likely give an additional increase in functional affinity (avidity), as demonstrated previously [38,39]. Furthermore, high selectivity of the Z HER2:342 and Z EGFR:1907 affibody molecules to their respective target protein has been shown in previous studies [30,39].…”

Section: Single and Dual Binding Biosensor Analysismentioning

confidence: 59%

“…Affibody molecules have been genetically fused to other proteins, e.g. ABD (albumin-binding domain) [36] or Fc fragment [37] or to another identical affibody molecule to increase the functional affinity (avidity) [38][39][40]. bs targeting has been achieved with an affibody-ABD fusion molecule, to allow binding to both the target protein and human serum albumin in order to obtain increased in vivo half-life [36] or to Fc to take advantage of Fc functions.…”

Section: Introductionmentioning

confidence: 99%

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Engineering and characterization of a bispecific HER2 × EGFR‐binding affibody molecule

Friedman

Lindström

Ekerljung

et al. 2009

Biotech and App Biochem

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Section: Single and Dual Binding Biosensor Analysissupporting

confidence: 86%

Section: Single and Dual Binding Biosensor Analysismentioning

confidence: 59%

Section: Introductionmentioning

confidence: 99%

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Engineering and characterization of a bispecific HER2 × EGFR‐binding affibody molecule

Friedman

Lindström

Ekerljung

et al. 2009

Biotech and App Biochem

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“…In Vivo Cellular and Molecular Imaging (ICMI) Laboratory, Vrije Universiteit Brussel (VUB), Brussels, Belgium; 2 Department of Molecular and Cellular Interactions, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Vrije Universiteit Brussel (VUB), Brussels, Belgium; 3 Nuclear Medicine Department, UZ Brussel, Brussels, Belgium; 4 Ablynx N.V., Zwijnaarde, Belgium; 5 Department for Molecular Biomedical Research, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Brussels, Belgium; 6 Department of Molecular Biology, Ghent University, Ghent, Belgium; 7 Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel (VUB), Brussels, Belgium…”

mentioning

confidence: 99%

“…As a result, the targeted tumors can be visualized only at several hours or even days after tracer injection (2,3). Radiolabeling of receptor ligands has also been reported (4)(5)(6). Blood clearance of these tracers is rapid, but the tumor uptake is only limited.…”

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confidence: 99%

Comparison of the Biodistribution and Tumor Targeting of Two ^99mTc-Labeled Anti-EGFR Nanobodies in Mice, Using Pinhole SPECT/Micro-CT

Gainkam¹,

et al. 2008

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Camelidae possess an unusual class of antibodies devoid of light chains. Nanobodies are intact antigen-binding fragments that are stable, easily generated against different targets, and fully functional. Their rapid clearance from the blood circulation favors their use as imaging agents. We compared the in vivo tumor uptake and biodistribution of 2 anti-epidermal growth factor receptor (anti-EGFR) Nanobodies, 99m Tc-7C12 and 99m Tc-7D12. Methods: Nanobodies were labeled via their hexahistidine tail with 99m Tc-tricarbonyl ( 99m Tc(CO) 3 ) generated from a kit. Mice bearing subcutaneous A431 (EGFR-positive) and R1M (EGFRnegative) xenografts were intravenously injected with 99m Tc-7C12 and 99m Tc-7D12 on separate days. Pinhole SPECT/ micro-CT images were acquired at 1 h after administration to assess noninvasively the biodistribution and tumor targeting of the labeled compounds. Pinhole SPECT and micro-CT images from the same mouse were automatically fused on the basis of a mathematic rigid-body-transformation algorithm using six 57 Co sources. Images were quantified, and tracer uptake was expressed as percentage injected activity per gram per cubic centimeter (%IA/cm 3 ) of tissue. Ex vivo biodistribution of mice bearing A431 injected with either 99m Tc-7C12 or 99m Tc-7D12 was also assessed; activity in the tumor and organs was recorded and expressed as percentage injected activity per gram (%IA/g). Results: Binding of both tracers was receptorspecific. Image analysis showed high and similar tumor uptake values for both 99m Tc-7C12 and 99m Tc-7D12 (4.55 6 0.24 %IA/ cm 3 and 4.62 6 0.36 %IA/cm 3 , respectively) in A431 xenografts, whereas the uptake in the negative tumor (R1M) was low (1.16 6 0.14 for 99m Tc-7C12 and 1.49 6 0.60 for 99m Tc-7D12). 99m Tc-7C12 showed significantly higher kidney uptake (63.48 6 2.36 vs. 56.25 6 2.46 %IA/cm 3 ) and lower liver uptake (2.55 6 0.26 vs. 4.88 6 0.86 %IA/cm 3 ) than did 99m Tc-7D12. The ex vivo analysis confirmed the image quantification with high tumor-tobackground ratio; however, 99m Tc-7C12 showed higher tumor uptake (9.11 6 1.12 %IA/g) than did 99m Tc-7D12 (6.09 6 0.77 %IA/g). 99m Tc-7D12 demonstrated significantly higher blood activity than did 99m Tc-7C12, but both showed short plasma half-lives (,10 min).Conclusion: The Nanobody fragments used here show high tumor uptake, low liver uptake, and rapid blood clearance. Nanobodies are promising probes for noninvasive radioimmunodetection of specific targets early after administration. On the basis of its favorable biodistribution, 99m Tc-7C12 was selected for further studies. Epi dermal growth factor receptor (EGFR or ErbB1) is a member of a receptor tyrosine kinase family together with Her-2-neu/ErbB2, HER-3/ErbB3, and HER-4/ErbB4. EGFR is implicated in many cellular processes such as proliferation, differentiation, and survival (1). Several reports have shown that EGFR signaling is abnormal in many tumors of epithelial origin, such as cancer of the breast, head and neck, prostate, lung, and skin. Aberrant signaling of...

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Metal Carbonyl–Gold Nanoparticle Conjugates for Live‐Cell SERS Imaging

Kong¹,

Lam²,

Goh³

et al. 2012

Angewandte Chemie

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The field of bioorganometallic chemistry, the junction of organometallic chemistry and biology, is coming of age. In particular, applications of metal carbonyl compounds in biological immunoassays (metallo-immunoassays), pharmaceuticals, CO therapy (CO-releasing molecules), and bioimaging have emerged. [1][2][3][4] One of the useful characteristics of metal carbonyl compounds is the strong CO stretching vibrations in the mid-IR region (1800-2200 cm À1 ); a region that is relatively free of interference from absorbance of biomolecules. This has been utilized in the development of carbonyl immunoassays, [1f] and more recently, in cell imaging, with detection techniques that include FTIR, Raman, and PTIR spectroscopy. [4] Although the first two detection methods can be carried out with readily available instrumentation, a major hindrance to IR detection for live-cell imaging is the interference from a strong absorption band of water at approximately 1600 cm À1 . Although Raman spectroscopy provides a far better spatial resolution with minimal interference from water, [5] it suffers from a low scattering cross section. [6] This necessitates a high concentration of the metal carbonyl based biotag, which poses the problem of undesired cytotoxicity and hence limits its potential in clinical applications. Furthermore, most metal carbonyl compounds have low solubility in water, thus requiring complicated bioconjugation to render them more water soluble or dispersible, and biocompatible. Therefore, there is a need to develop a highly sensitive, targeted, and low-toxicity metal carbonyl based biotag toward eventual biological applications.One promising approach to signal enhancement is surface-enhanced Raman spectroscopy (SERS); the Raman signals of molecules on colloidal gold or silver nanoparticles can be enhanced by several orders of magnitude (typically 10 6 to 10 14 ) as a result of the strong surface plasmon resonance of the nanostructured surface. This spectroscopic method has been successfully adapted to chemical sensing applications, at lower concentrations but with better detection limits, [7] and is exemplified by its use in DNA detection, [8] cancer diagnosis, [9] and detection of cellular molecules. [10] The possibility that SERS may also be used to enhance the CO stretching vibration signal of metal carbonyl compounds is attractive. Furthermore, detection by Raman microscopy will afford spatial resolution into the submicron range.The use of organometallic osmium carbonyl clusters (OM) has been demonstrated earlier; [4a] they have the advantages of being oxygen-and moisture-stable, and are very robust in comparison with other organometallic compounds. The preparation and mode of interaction of osmium carbonyl clusters with the surface of gold nanoparticles (NPs) have also been reported recently. [11] We thus envisage that the binding of an organometallic osmium carbonyl cluster, Os 3 (CO) 10 (m-H) 2 , onto the surface of gold nanoparticles (henceforth designated as "OM-NP") may significantly enhance the i...

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Phage display selection of Affibody molecules with specific binding to the extracellular domain of the epidermal growth factor receptor

Cited by 107 publications

References 53 publications

Engineering and characterization of a bispecific HER2 × EGFR‐binding affibody molecule

Engineering and characterization of a bispecific HER2 × EGFR‐binding affibody molecule

Comparison of the Biodistribution and Tumor Targeting of Two ^99mTc-Labeled Anti-EGFR Nanobodies in Mice, Using Pinhole SPECT/Micro-CT

Metal Carbonyl–Gold Nanoparticle Conjugates for Live‐Cell SERS Imaging

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Phage display selection of Affibody molecules with specific binding to the extracellular domain of the epidermal growth factor receptor

Cited by 107 publications

References 53 publications

Engineering and characterization of a bispecific HER2 × EGFR‐binding affibody molecule

Engineering and characterization of a bispecific HER2 × EGFR‐binding affibody molecule

Comparison of the Biodistribution and Tumor Targeting of Two 99mTc-Labeled Anti-EGFR Nanobodies in Mice, Using Pinhole SPECT/Micro-CT

Metal Carbonyl–Gold Nanoparticle Conjugates for Live‐Cell SERS Imaging

Contact Info

Product

Resources

About

Comparison of the Biodistribution and Tumor Targeting of Two ^99mTc-Labeled Anti-EGFR Nanobodies in Mice, Using Pinhole SPECT/Micro-CT