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首页> 外文期刊>Scientific reports. >A standardized quantitative method for detecting remnant alpha-Gal antigen in animal tissues or animal tissue-derived biomaterials and its application
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A standardized quantitative method for detecting remnant alpha-Gal antigen in animal tissues or animal tissue-derived biomaterials and its application

机译：检测动物组织或动物组织来源的生物材料中残留α-Gal抗原的标准化定量方法及其应用

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摘要

Alpha-Gal (Gal) epitopes present in animal tissues are known to be the key xenoantigens that elicit xenorejection. However, a standardized method to determine Gal epitope in animal tissue-derived biomaterials does not exist. Herein, a standardized method for quantitative detection of Gal antigen was established based on an ELISA inhibition assay with Gal antibody. In this method, the key optimized experimental conditions were: (1) Gal-antigen positive and negative reference materials were developed, and used as positive and negative control in the test system, respectively; (2) A mixture of artificial Gal-BSA antigen plus Gal-negative matrix was used as the calibration standard sample, making it has similar composition with test sample; and (3) The lysis buffer was combined with the homogenate to expose the Gal antigen as much as possible. The results from validation and application experiments showed that the standardized method had good reproducibility (RSD?=?12.48%), and the lower detection limit (LDL) is ~7.1?×?1011?Gal epitopes/reaction. This method has been further developed into a detection Kit (Meitan 70101, China), and it has been developed as a standard method for detecting remnant immunogen of animal tissue derived medical devices, and as the industry standard has been released in China. (YY/T 1561–2017).
机译：已知存在于动物组织中的Al-Gal（Gal）表位是引起异种注射的关键异种抗原。但是，不存在确定动物组织来源的生物材料中Gal表位的标准化方法。在此，基于使用Gal抗体的ELISA抑制试验，建立了定量检测Gal抗原的标准化方法。该方法优化的关键实验条件为：（1）开发了Gal抗原阳性和阴性参考物质，分别用作测试系统的阳性和阴性对照。 （2）将人工Gal-BSA抗原加Gal阴性基质的混合物用作校正标准样品，使其组成与测试样品相似。 （3）将裂解缓冲液与匀浆组合以尽可能暴露Gal抗原。验证和应用实验的结果表明，该标准化方法具有良好的重现性（RSD≥12.48％），检测下限（LDL）为〜7.1××1011 -Gal表位/反应。该方法已被进一步开发为检测试剂盒（中国棉兰70101），已被开发为检测动物组织来源的医疗器械残留免疫原的标准方法，并已在中国发布行业标准。 （YY / T 1561–2017）。

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