Patent ID: 11898187
Assignee: NORTHWESTERN UNIVERSITY
Field: Measurement (Instruments)
Classification: CPC C  G | IPC C  G

Claim 0:
1. A method for synthesizing a glycoprotein or recombinant glycoprotein, the method comprising:
(a) expressing in a cell or in a cell-free protein synthesis (CFPS) reaction, a polypeptide comprising the amino acid sequence of a target protein which includes naturally an amino acid motif that includes an asparagine that is glycosylated by N-glycosyltransferase from Actinobacillus pleuropneumoniae (ApNGT) or that has been modified to include a heterologous amino acid motif that includes an asparagine that is glycosylated by ApNGT, the amino acid motif or heterologous amino acid motif comprises X−2-X−1-N-X+1-S/T-X+3, wherein X−2 is selected from Gly, Asn, and Tyr; X−1 is selected from Gly and Ala; X+1 is selected from Trp, Val, His, Ala, and Ile; and X+3 is selected from Thr, Met, and Phe;
(b) expressing ApNGT in a cell or in a CFPS reaction; and
(c) reacting the polypeptide and ApNGT in the presence of a sugar donor, wherein ApNGT glycosylates the amino acid motif or heterologous amino acid motif of the polypeptide with the sugar of the sugar donor to synthesize the glycoprotein or recombinant glycoprotein.