title: Kit formulation for 99mTc-labeling of recombinant anti-HER2 Affibody molecules with a C-terminally engineered cysteine.
abstract: BACKGROUND
  Molecular imaging of human epidermal growth factor receptor type 2 (HER2)-expression in malignant tumors provides potentially important information for patient management. Affibody molecules have shown to be suitable tracers for imaging applications using single photon emission computed tomography or positron emission tomography. Results from an earlier evaluation of the application of site-specific (99m)Tc-labeling of the Affibody molecule, Z(HER2:2395)-C, were favorable.
  
  
  METHODS
  As a preparation for clinical application of this tracer, we have developed and evaluated a robust single-vial freeze-dried kit, allowing labeling of the Affibody molecule, Z(HER2:2395)-C, with (99m)Tc.
  
  
  RESULTS
  The composition of the kit [containing glucoheptonate, EDTA and tin(II)-chloride], as well as the protein amount and the pertechnetate volume were optimized for a high labeling yield (>90%) and minimal presence of reduced hydrolyzed technetium colloids (<1%). The specificity to HER2 receptors, the binding competence and the stability in phosphate-buffered saline and murine serum were verified in vitro. The shelf-life was also evaluated in vitro, showing no reduction in labeling yield or binding capacity to HER2-expressing cells after over 400 days of storage of the single-vial freeze-dried kit.
  
  
  CONCLUSIONS
  Z(HER2:2395)-C labeled with (99m)Tc using the lyophilized kit was stable and resulted in a favorable biodistribution in an in vivo evaluation in normal Naval Medical Research Institute mice.
authors.affiliation: Division of Nuclear Medicine, Department of Medical Sciences, Uppsala University, SE-751 85, Uppsala, Sweden. sara.ahlgren@medsci.uu.se
authors.forename: Sara
authors.initials: S
authors.lastname: Ahlgren
authors.forename: Kristofer
authors.initials: K
authors.lastname: Andersson
authors.forename: Vladimir
authors.initials: V
authors.lastname: Tolmachev
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor; D003545:Cysteine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004492:Edetic Acid / Q000737:chemistry; D005260:Female; D005612:Freeze Drying; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D051379:Mice; D015609:Organotechnetium Compounds / Q000737:chemistry*; D015202:Protein Engineering; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D013670:Sodium Pertechnetate Tc 99m / Q000737:chemistry; D013400:Sugar Acids / Q000737:chemistry; D013997:Time Factors; D017971:Tin Compounds / Q000737:chemistry
chemical_list: D000906:Antibodies; D003545:Cysteine; D004492:Edetic Acid; D015609:Organotechnetium Compounds; D018719:Erb-b2 Receptor Tyrosine Kinases; D011994:Recombinant Proteins; D013670:Sodium Pertechnetate Tc 99m; D013400:Sugar Acids; D017971:Tin Compounds; C023599:stannous chloride; C007852:alpha-glucoheptonic acid