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Nordberg E , Friedman M , Gostring L , Adams GP , Brismar H , Nilsson FY , Stahl S , Glimelius B , Carlsson J
Cellular studies of binding, internalization and retention of a radiolabeled EGFR-binding affibody molecule
Nuclear Medicine and Biology. 2007 Aug;34(6) :609-618
AbstractIntroduction: The cellular binding and processing of an epidermal growth factor receptor (EGFR) targeting affibody molecule, (Z(EGFR:955))(2), was studied. This new and small molecule is aimed for applications in nuclear medicine. The natural ligand epidermal growth factor (EGF) and the antibody cetuximab were studied for comparison. Methods: All experiments were made with cultured A431 squamous carcinoma cells. Receptor specificity, binding time patterns, retention and preliminary receptor binding site localization studies were all made after (125) I, labeling. Internalization was studied using Oregon Green 488, Alexa Fluor 488 and CypHer5E markers. Results: [(125) I](Z(EGFR:955))(2) and [(125) I]cetuximab gave a maximum cellular uptake of I-125 within 4 to 8 h of incubation, while [(125) I]EGF gave a maximum uptake already after 2 h. The retention studies showed that the cell-associated fraction of 125 1 after 48 It of incubation was similar to 20% when delivered as [(125) I](Z(FGFR:955))(2) and similar to 25% when delivered as [I-125] cetuximab. [(125) I]EGF-mediated delivery gave a faster (125) I release, where almost all cell-associated radioactivity had disappeared within 24 It. All three substances were internalized as demonstrated with confocal microscopy. Competitive binding studies showed that both EGF and cetuximab inhibited binding Of (Z(EGFR:955))(2) and indicated that the three substances competed for an overlapping binding site. Conclusion: The results gave information on cellular processing of radionuclides when delivered with (Z(EGFR:955))(2) in comparison to delivery with EGF and cetuximab. Competition assays suggested that [I-125](Z(EGFR:955))(2) bind to Domain III of EGFR. The affibody molecule (Z(EGFR:955))(2) can be a candidate for EGFR imaging applications in nuclear medicine. (c) 2007 Elsevier Inc. All rights reserved.
NotesISI Document Delivery No.: 206AW Times Cited: 0 Cited Reference Count: 30 Cited References: BASELGA J, 2005, J CLIN ONCOL, V23, P2445 BAUM RP, 2006, J NUCL MED, V47, P108 BURGESS AW, 2003, MOL CELL, V12, P541 CARLSSON J, 2003, RADIOTHER ONCOL, V66, P107 CARLSSON J, 2006, CURR CANC THER REV, V2, P293 CHO HS, 2003, NATURE, V421, P756 CITRI A, 2003, EXP CELL RES, V284, P54 CITRI A, 2006, NAT REV MOL CELL BIO, V7, P505 EARP HS, 1995, BREAST CANCER RES TR, V35, P115 FERGUSON KM, 2003, MOL CELL, V11, P507 FRIEDMAN M, 2007, PROTEIN ENG DES SEL, V20, P189 GARRETT TPJ, 2003, MOL CELL, V11, P495 GOLDSTEIN NI, 1995, CLIN CANCER RES, V1, P1311 HARARI PM, 2004, ENDOCR-RELAT CANCER, V11, P689 JARAMILLO ML, 2006, EXP CELL RES, V312, P2778 LI SQ, 2005, CANCER CELL, V7, P301 MARSHALL J, 2006, CANCER, V107, P1207 MENDELSOHN J, 2003, J CLIN ONCOL, V21, P2787 NORD K, 1997, NAT BIOTECHNOL, V15, P772 NORMANNO N, 2006, GENE, V366, P2 QIAN XL, 1994, P NATL ACAD SCI USA, V91, P1500 STEFAN S, 1999, ENCY BIOPROCESS TECH STEFFEN AC, 2005, CANCER BIOTHER RADIO, V20, P239 STEFFEN AC, 2006, EUR J NUCL MED MOL I, V33, P631 SUNDBERG AL, 2003, NUCL MED BIOL, V30, P303 TOLMACHEV V, 2006, J NUCL MED, V47, P846 WARREN CM, 2006, CELL SIGNAL, V18, P923 WILEY HS, 2003, EXP CELL RES, V284, P78 YARDEN Y, 1987, BIOCHEMISTRY-US, V26, P1443 YARDEN Y, 2001, NAT REV MOL CELL BIO, V2, P127