Preparation and purification of an end to end coupled mEGF-dextran conjugate.
The amino terminus of mouse epidermal growth factor (mEGF) was coupled directly to the aldehyde end of dextran through a reductive amination procedure. The highest coupling efficiency was approximately 80% and could be reached after approximately 24 h of reaction time at pH 8. Gel filtration on Sephadex G-50 Fine removed free mEGF from the conjugate. Preparative polyacrylamide gel electrophoresis was used to separate the conjugate from excess noncharged dextran. The conjugate bound specifically to the EGF receptor on cultured glioma cells as shown in displacement tests with free mEGF. The conjugate was stable in the pH interval 4-9, in 2 M sodium chloride, in 7 M urea, and in human serum and could still bind to the EGF receptor after such treatments. The conjugates are candidates for targeted nuclide therapy.
Zhao
Q
Q
Department of Diagnostic Radiology, Uppsala University, Sweden. Qinghai.Zhao@bms.uu.se
Gottschalk
I
I
Carlsson
J
J
Arvidsson
L E
LE
Oscarsson
S
S
Medin
A
A
Ersson
B
B
Janson
J C
JC
D000586:Amination; D000818:Animals; D003911:Dextrans / Q000138:chemical synthesis* / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004815:Epidermal Growth Factor / Q000138:chemical synthesis* / Q000302:isolation & purification* / Q000378:metabolism; D005910:Glioma / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007457:Iodine Radioisotopes; D051379:Mice; D010084:Oxidation-Reduction; D014407:Tumor Cells, Cultured
D003911:Dextrans; D007457:Iodine Radioisotopes; C067291:epidermal growth factor-dextran conjugate; D004815:Epidermal Growth Factor; C025614:sephadex