Method of radio-labelling biomolecules
A radioactive labeling, biomolecule technology, applied in the directions of radioactive physical shape, radioactive carrier, radioactive preparation in vivo, etc., can solve the problems of non-imaging agent, low tumor ratio, low activity, etc.
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Embodiment 1
[0097] Apo-lactotransferrin (aLf) was pre-incubated with 35 [mu]M of 2-mercaptoethanol overnight at 4[deg.]C prior to use and then labeled with 1.6 [mu]M of labeled protein at 7% efficiency. Labeled proteins were repeatedly washed and 92% of the label was still attached after 60 min incubation in PBS. The results are shown in Table 3.
Embodiment 2
[0099] Apotransferrin (aTf) was pre-incubated overnight at 4°C with various concentrations of 2.6 μM, 12 μM and 26 μM 2-mercaptoethanol (2-ME) before use, yielding 8%, 25% and 60% labeling, respectively efficiency. Repeating the lowest concentration of 2-ME gave labeling efficiencies of 7% and 8.5%. After washing and 90 min incubation at 37°C in PBS, 97% of the highest labeled concentration of aTf remained on the protein. The results are shown in Table 3.
Embodiment 3
[0101] DTPA is an exemplary chelating moiety in the prior art and forms stable chelates with various metals. The amount of labeling on aTf decreased by about 10% after 2 h incubation in the presence of DTPA, from 97% to 87%, indicating removal of weakly bound radionuclides from biomolecules.
[0102] Incubation of aTf in acidic conditions also resulted in a reduction in the amount of bound radionuclides. The amount of label on the protein decreased by about 23% after 2 hours of incubation at pH 5, from 97% to 75%, see Table 4.
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