Ngwaahịa

Nnwale chromatography mmiri na-arụ ọrụ ọhụrụ maka glycosyltransferases dabere na mwepu ya na anthranilic acid na nchọpụta fluorescence.

Emepụtara nyocha site na iji kemịkalụ akara pụrụ iche nke 2-aminobenzoic acid (2AA; anthranilic acid, AA) maka ịlele ọrụ nke ma β1-4 galactosyltransferase (GalT-1) na α2-6 sialyltransferase (ST-6) site na mmiri mmiri na-arụ ọrụ dị elu. chromatography (HPLC) na nchọpụta fluorescence (Anumula KR. 2006. Ọganihu na fluorescence derivatization ụzọ maka elu-arụmọrụ mmiri chromatographic analysis nke glycoprotein carbohydrates. Anal Biochem. 350:1-23).A na-eji N-Acetylglucosamine (GlcNAc) na N-acetyllactosamine dị ka ndị na-anabata ya na uridine diphosphate (UDP) -galactose na cytidine monophosphate (CMP) -N-acetylneuraminic acid (NANA) dị ka ndị na-enye onyinye maka GalT-1 na ST-6, n'otu n'otu.Edebere ngwaahịa enzymatic n'ọnọdụ ya na AA wee kewapụ ya na ihe ndị dị na kọlụm TSKgel Amide 80 site na iji ọnọdụ-usoro nkịtị.Ekpebiri nkeji enzyme site na mpaghara kacha elu site na ntụnyere ụkpụrụ Gal-β1-4GlcNAc na NANA-α2-6 Gal-β1-4GlcNAc.Linearity (oge na ntinye enzyme), nkenke (intra- na interassay) na reproducibility maka nyocha ndị e guzobere.Achọpụtara nyocha ndị ahụ ka ọ bara uru na nyochaa ọrụ enzyme n'oge ikewapụ na ịdị ọcha.Nnwale ndị ahụ nwere mmetụta nke ukwuu ma mee ha nhata ma ọ bụ ka mma karịa nha dabere na redioaktivu ọdịnala.A pụkwara iji usoro nyocha ahụ maka ịlele ọrụ nke nnyefe ndị ọzọ, ma ọ bụrụhaala na ndị na-anabata carbohydrate nwere njedebe mbelata maka ịde aha.Emepụtara nyocha maka ndị na-anabata glycoprotein site na iji IgG.E mepụtara usoro profaịlụ HPLC dị mkpirikpi maka nkewa nke IgG glycans (biantennary G0, G1, G2, mono- na diialylated), nke mere ka mkpebi nke GalT-1 na ST-6 dị ngwa ngwa.Ọzọkwa, usoro profaịlụ a kwesịrị ịba uru maka nyochaa mgbanwe na IgG glycans na ntọala ụlọ ọgwụ.