The resuspended cells can be homogenized under chilled conditions and centrifuged (Sorvall, Thermofisher)
The gel purified His6-AV-Ubiquitin-semapeptide (nucleotide sequence) encoding gene PCR product and pET 24 plasmid vector was restriction digested (double digestion) with Ndel (NEB) + Xhol (NEB) at 37 C
Solid phase synthesis: - The synthesis of semaglutide usually starts with solid phase synthesis
Two carboxyl groups on a side chain disclosed by patent CN111378028A are naked and are not protected by a protective group, the purity of an intermediate obtained in the subsequent coupling reaction with an Arg34GLP-1 (9-37) main peptide chain is not high, only 67.5 percent is obtained, the yield is seriously reduced, and the pH value in the reaction has a large influence on the side chain modification, so that various byproducts which are difficult to remove are easily generated, such as a double-modified impurity byproduct with both the 26 th position and the end position of the main peptide chain modified and a byproduct with the end position of the main peptide chain modified
This makes the overall treatment with semaglutide more comfortable and tolerable, ensuring better adherence and more consistent results for patients
