Lihlahisoa

Teko e ncha e sebetsang hantle haholo ea mokelikeli oa chromatography bakeng sa glycosyltransferase e ipapisitseng le ho tsoa ho anthranilic acid le tlhahlobo ea fluorescence.

Litlhahlobo li entsoe ho sebelisoa k'hemistri e ikhethang ea 2-aminobenzoic acid (2AA; anthranilic acid, AA) bakeng sa ho lekanya mesebetsi ea bobeli ba β1-4 galactosyltransferase (GalT-1) le α2-6 sialyltransferase (ST-6) ka mokelikeli o sebetsang hantle. chromatography (HPLC) e nang le ho lemoha ha fluorescence (Anumula KR. 2006. Tsoelo-pele ea mekhoa ea fluorescence derivatization bakeng sa tlhahlobo e phahameng ea ts'ebetso ea mokelikeli oa chromatographic ea lik'habohaedreite tsa glycoprotein. Anal Biochem. 350: 1-23).N-Acetylglucosamine (GlcNAc) le N-acetyllactosamine li ile tsa sebelisoa e le ba amohelang le uridine diphosphate (UDP) -galactose le cytidine monophosphate (CMP) -N-acetylneuraminic acid (NANA) e le bafani ba GalT-1 le ST-6, ka ho latellana.Lihlahisoa tsa enzymatic li ne li ngotsoe sebakeng se nang le AA 'me li arotsoe ho tloha ho li-substrates ho TSKgel Amide 80 kholomo ho sebelisoa maemo a tloaelehileng.Likarolo tsa enzyme li ile tsa khethoa ho tloha libakeng tse phahameng ka ho bapisoa le litekanyetso tse nkiloeng ka nako e tšoanang Gal-β1-4GlcNAc le NANA-α2-6 Gal-β1-4GlcNAc.Linearity (nako le mahloriso a enzyme), ho nepahala (intra- and interassay) le reproducibility bakeng sa litlhahlobo li ile tsa thehoa.Litlhahlobo li fumanoe li le molemo ho lekola mesebetsi ea enzyme nakong ea ho itšehla thajana le tlhoekiso.Litlhahlobo li ne li le hlokolosi haholo 'me li etsoa tse lekanang kapa li betere ho feta litekanyo tse tloaelehileng tse thehiloeng ho tsoekere ea radioactive.Sebopeho sa assay se ka boela sa sebelisoa ho lekanya tšebetso ea li-transferase tse ling, ha feela ba amohelang lik'habohaedreite ba na le qetello e fokolang bakeng sa ho ngola.Teko ea ba amohelang glycoprotein e entsoe ka IgG.Mokhoa o mokhuts'oane oa ho etsa profil ea HPLC o ile oa ntlafatsoa bakeng sa karohano ea IgG glycans (biantennary G0, G1, G2, mono- le disialylated), e ileng ea thusa ho etsa qeto ea mesebetsi ea GalT-1 le ST-6 ka potlako.Ho feta moo, mokhoa ona oa ho etsa profil e lokela ho ba molemo bakeng sa ho lekola liphetoho tsa IgG glycans maemong a kliniki.