The invention discloses a synthesis method of a uniform sialylated AFP-L3
antigen, serum polyclonal
antibody preparation and application thereof, and belongs to the field of
immunology, the uniform sialylated AFP-L3
antigen comprises a sialylated AFP-L3
glycopeptide antigen or a sialylated AFP-L3
glycoprotein antigen, and the synthesis method comprises the following steps: transferring a
carbohydrate chain to a starting substrate by using Endo-M N175Q transglycosylation reaction to obtain an intermediate I; wherein the starting substrate comprises a
glycopeptide Fmoc-ATKVN (GlcNAc) FTEAQKAALDV-NH2 or an AFP (Alpha
Fetal Protein)
protein which is expressed in HEK293T and is subjected to
hydrolysis treatment; modifying the intermediate I through
fucose transferase FuT8 to obtain an intermediate II; the intermediate II is modified and extended through
galactose transferase GalT and
sialyltransferase SiaT, and an intermediate III or a sialylated AFP-L3
glycoprotein antigen is obtained; the sialylated AFP-L3
glycopeptide antigen is obtained through the steps that the sialylated AFP-L3 glycopeptide antigen is obtained, the intermediate III is subjected to Fmoc removal treatment and then conjugated with bis-
succinimide glutaric acid ester and
hemocyanin, and the serum polyclonal
antibody generated based on the antigen has
high titer and specificity.