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    • 2. 发明公开
    • METHOD FOR DETERMINING SITE HAVING N-LINKED SUGAR CHAIN ADDED THERETO OR PROPORTION OF SAID ADDITION
    • VERFAHREN ZUR BESTIMMUNG EINER STELLE麻省理工N-VERBUNDENER ZUCKERKETTE,DIE DAZU HINZUGEGEBEN WURDE,ODER DER比例DER BESAGTEN ZUGABE
    • EP3088531A4
    • 2017-06-14
    • EP14874116
    • 2014-12-17
    • TOSOH CORP
    • NAKATA DAISUKE
    • C12Q1/34C12N9/16C12Q1/37G01N27/62
    • C12Q1/34C07K14/57C12N9/80C12Y305/01052G01N33/6866G01N2333/57G01N2333/98G01N2440/38
    • A method for detecting a site which can be modified with an N-linked glycan chain and to which an N-linked glycan chain is actually linked in a glycoprotein; and a method for determining the state of an N-linked glycan chain addition at the site are provided. A glycoprotein having an N-linked glycan chain linked thereto is subjected to an N-linked glycan chain removal treatment with a peptide N-glycanase, subsequently a site capable of being modified with an N-linked glycan chain, in which an Asn residue has been changed to an Asp residue by the action of the peptide N-glycanase, is treated with an endo-type peptidase capable of recognizing an Asp residue to thereby produce peptide fragments, and subsequently the mass of the fragments is detected. In this manner, a site which can be modified with an N-linked glycan chain and to which an N-linked glycan chain is actually linked can be detected. Furthermore, the proportion or state of of the N-linked glycan chain addition at the site can be determined from the intensity of a signal generated upon the detection.
    • 用于检测可以用N-连接的聚糖链修饰并且N-连接的聚糖链实际上连接在糖蛋白上的位点的方法; 并提供了用于确定在该位点处N-连接的聚糖链添加状态的方法。 将具有与其连接的N-连接的聚糖链的糖蛋白用肽N-聚糖酶进行N-连接的聚糖链去除处理,随后用能够用N-连接的聚糖链修饰的位点进行N-连接的聚糖链去除处理,其中Asn残基具有 通过肽N-聚糖酶的作用改变为Asp残基,用能够识别Asp残基的内切型肽酶处理,从而产生肽片段,随后检测片段的质量。 以这种方式,可以检测可以用N-连接的聚糖链修饰并且N-连接的聚糖链实际连接到的位点。 此外,可以从检测时产生的信号的强度确定在该位点的N-连接的聚糖链添加的比例或状态。