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The preparation method according to claim 1 or 2, wherein the coupling in step 2 is performed by condensing by DIC+A or B+A+C method until the resin is detected to be transparent with ninhydrin, wherein A is chosen from HOBt or HOAt, B is chosen from HBTU, HATU, TBTU, PyAOP, or PyBOP, C is chosen from DIPEA or TMP
Cooling to 0 , stirring for 30min, adding 0.22g of HOSu, stirring and reacting for 12 hours at the temperature, filtering, collecting filtrate, and spin-drying the filtrate to obtain crude Fmoc-His-Aib-OSu
200 g Fmoc-Gly-Wang Resin (21.6 mmol) with a degree of substitution of 0.108 mmol/g was weighed and swelled for 30 min with DMF, followed by sequentially coupling according to the amino acid sequence of Semaglutide with 2-5 equivalents of amino acid feeds and condensing with DIC+A or B+A+C, until the resin was detected to be transparent with ninhydrin, wherein A was HOBt or HOAt
The transformed cells were grown on LB agar plate + Kanamycin at 37 C
