Nā huahana

ʻO ka hōʻike chromatography wai hou kiʻekiʻe no nā glycosyltransferases e pili ana i ka derivatization me ka anthranilic acid a me ka ʻike fluorescence.

Ua hoʻomohala ʻia nā hoʻokolohua me ka hoʻohana ʻana i ka kemika lepili kūʻokoʻa o 2-aminobenzoic acid (2AA; anthranilic acid, AA) no ke ana ʻana i nā hana o nā β1-4 galactosyltransferase (GalT-1) a me α2-6 sialyltransferase (ST-6) e ka wai hana kiʻekiʻe. chromatography (HPLC) me ka ʻike fluorescence (Anumula KR. 2006. ʻO nā holomua i nā ʻano derivatization fluorescence no ka hana kiʻekiʻe kiʻekiʻe o ka wai chromatographic analysis o glycoprotein carbohydrates. Anal Biochem. 350:1-23).Ua hoʻohana ʻia ʻo N-Acetylglucosamine (GlcNAc) a me N-acetyllactosamine ma ke ʻano he ʻae a me ka uridine diphosphate (UDP) -galactose a me cytidine monophosphate (CMP) -N-acetylneuraminic acid (NANA) ma ke ʻano he mau mea hāʻawi no GalT-1 a me ST-6.Ua hōʻailona ʻia nā huahana Enzymatic ma kahi me AA a ua hoʻokaʻawale ʻia mai nā substrates ma ke kolamu TSKgel Amide 80 me ka hoʻohana ʻana i nā kūlana maʻamau.Hoʻoholo ʻia nā ʻāpana enzyme mai nā wahi kiʻekiʻe ma ka hoʻohālikelike ʻana me nā maʻamau i hoʻohālikelike ʻia Gal-β1-4GlcNAc a me NANA-α2-6 Gal-β1-4GlcNAc.Ua hoʻokumu ʻia ka linearity (manawa a me ka enzyme concentration), precision (intra- and interassay) a me reproducibility no nā assays.Ua ʻike ʻia nā hoʻokolohua he mea pono i ka nānā ʻana i nā hana enzyme i ka wā kaʻawale a me ka hoʻomaʻemaʻe.He koʻikoʻi koʻikoʻi ka hoʻāʻo ʻana a ua like a ʻoi aku ka maikaʻi ma mua o nā ana radioactive maʻamau.Hiki ke hoʻohana ʻia ke ʻano hoʻāʻo no ke ana ʻana i ka hana o nā hoʻololi ʻē aʻe, inā i loaʻa i ka poʻe e ʻae ana i ka carbohydrate kahi hopena hōʻemi no ka lepili.Hoʻokumu ʻia kahi hoʻokolohua no nā mea ʻae glycoprotein me ka hoʻohana ʻana iā IgG.Ua hoʻomohala ʻia kahi ʻano hana profiling HPLC pōkole no ka hoʻokaʻawale ʻana i nā glycans IgG (bintennary G0, G1, G2, mono- a me disialylated), i maʻalahi i ka hoʻoholo ʻana o nā hana GalT-1 a me ST-6 ma ke ʻano wikiwiki.Eia kekahi, pono e hoʻohana pono kēia ʻano hana no ka nānā ʻana i nā loli i ka IgG glycans i nā hoʻonohonoho maʻi.