Once I know how the NHS / EDC works.
However, the subject I am going to experiment with is the amino-modified oligonucleotide (C6, 5 ') ... the protocol is related to the protein ... (I have not yet figured out where to conjugate, but maybe I think I will conjugate in polymer, PEG.)
So I would like to ask something.
1. When conjugating an oligonucleotide with an amine group, keep the pH between 8.5 and 9.5. Is this the same when NHS / EDC reaction is performed? The reason why the pH is kept as such is that the free aliphatic amine is lowered when the pH is lower than 8.
2. Does the phosphate in the backbone of the oligonucleotide affect the reaction?
3. Does activation proceed separately before processing EDC / NHS? Or is it activating while inserting NHS / EDC?
4. Are there any other precautions besides keeping the pH between 8.5 and 9.5? (Eg sample after processing must be refrigerated)
Hi Kyungsun!
Try to perform the reaction in an organic solvent (DFM, DMSO, DMAc, DCM) with a small amount of DIPEA or TEA. If your compounds are in the water, just lionize them and dissolve in an organic solvent.
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thx for your answer.
Is it OK to use EDC instead of DCC? And, I am trying to conjugate PEG-COOH with amino-modified oligonucleotides. Is this a junction with this protocol?
EDC is water soluble, DCC isn't. EDC is also soluble in organic solvents e.g. DCM, THF, DMF, EtOH, and MeOH. The organic environment will be good for your case. Use DCM or MeOH, because these are easy to evaporate. Yes, this protocol is appropriate, but you can use more organic base.
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