The interaction between IgA tissue transglutaminase (tTG) antibodies (Abs) and 35S-labelled tTG produced in a transcription/translation (TnT) system with various amino acid (aa) deletions has been studied. These experiments showed that the tTG N-terminal aa 1-89 were important for tTG Ab binding in all 15 coeliac disease sera studied and the central residues (aa 401-491) were important for binding of tTG Abs in all but one sera. The contribution of C-terminal residues to tTG Ab binding varied in different coeliac sera but overall was less than the contributions of the N terminal and central regions. Mouse monoclonal antibodies (MAbs) to tTG were produced and the tTG aa sequences recognised by the MAbs determined using modified 35S-labelled tTG proteins. Analysis of the inhibiting effects of patient sera tTG Ab on binding of tTG MAbs to tTG confirmed the importance of the N-terminal and central regions of tTG in forming serum tTG Ab binding sites. Recombinant human tTG was expressed in yeast and purified to better than 95% homogeneity using MAb affinity chromatography as a final purification step. This material was highly suitable for use in an ELISA for tTGAb.

中文翻译：

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乳糜泻中组织转谷氨酰胺酶抗体识别的表位。

研究了IgA组织转谷氨酰胺酶（tTG）抗体（Abs）与带有各种氨基酸（aa）缺失的转录/翻译（TnT）系统中产生的35S标记的tTG之间的相互作用。这些实验表明，tTG N-末端aa 1-89对所有研究的15种腹腔疾病血清中的tTG Ab结合都很重要，而中央残基（aa 401-491）对于除一种血清之外的所有血清中的tTG Ab都很重要。C末端残基对tTG Ab结合的贡献在不同的腹腔血清中有所不同，但总体上小于N末端和中部区域的贡献。产生了针对tTG的小鼠单克隆抗体（MAbs），并使用修饰的35S标记的tTG蛋白确定了单克隆抗体识别的tTG aa序列。分析患者血清tTG Ab对tTG MAb与tTG结合的抑制作用，证实了tTG的N末端和中央区域在形成血清tTG Ab结合位点方面的重要性。重组人tTG在酵母中表达，并使用MAb亲和色谱作为最终纯化步骤纯化至优于95％的同质性。该材料非常适合用于tTGAb的ELISA。