CN114736297A - 一种高特异性的心肌肌钙蛋白i抗体及其制备方法和用途 - Google Patents
一种高特异性的心肌肌钙蛋白i抗体及其制备方法和用途 Download PDFInfo
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Abstract
本发明涉及心肌肌钙蛋白I抗体技术领域，尤其为一种高特异性的心肌肌钙蛋白I抗体及其制备方法和用途，心肌肌钙蛋白I抗体包括210个氨基酸组成，分子量23.9，等电点9.87，是一种富含α螺旋的蛋白质，有5个螺旋区域，呈伸展式构型，心肌肌钙蛋白I抗体(cTnl)为鼠单抗，体外纯化细胞培养上清获得，用于免疫发光和化学发光，本发明中，心肌肌钙蛋白I抗体(cTnl)抗体采用体外纯化细胞培养上清得到，与腹水来源的心肌肌钙蛋白I抗体(cTnl)相比，细胞体外培养生产的抗体避免了腹水中其它鼠源成分的干扰，产品质量更好，同时批次间差异更小，更稳定。
Description
技术领域
本发明涉及心肌肌钙蛋白I抗体技术领域，具体为一种高特异性的心肌肌钙蛋白I抗体及其制备方法和用途。
背景技术
由于心肌肌钙蛋白I抗体(cTnl)具有组织特异性强、宽阔的诊断窗口期、明确的诊断界限、快速的测定方法、血液中出现早等优点，在临床上以得到广泛应用，心肌肌钙蛋白I抗体(cTnl)是心肌细胞损伤最敏感和最特异性的血清标志物之一,加之心肌肌钙蛋白I抗体(cTnl)具有诊断阈值明确、窗口期宽以及检测快速等优点，已逐渐判断AMI患者心肌细胞损伤的主要生化指标，因此，对一种高特异性的心肌肌钙蛋白I抗体及其制备方法和用途的需求日益增长。
目前市场上存在的心肌肌钙蛋白I抗体(cTnl)来源腹水，存在其它鼠源成分的干扰，产品质量一般，而且批次间差异较大，稳定性差，因此，针对上述问题提出一种高特异性的心肌肌钙蛋白I抗体及其制备方法和用途。
发明内容
本发明的目的在于提供一种高特异性的心肌肌钙蛋白I抗体及其制备方法和用途，以解决上述背景技术中提出的问题。
为实现上述目的，本发明提供如下技术方案：
一种高特异性的心肌肌钙蛋白I抗体及其制备方法和用途，心肌肌钙蛋白I抗体包括210个氨基酸组成，分子量23.9，等电点9.87，是一种富含α螺旋的蛋白质，有5个螺旋区域，呈伸展式构型，心肌肌钙蛋白I抗体(cTnl)为鼠单抗，体外纯化细胞培养上清获得，用于免疫发光和化学发光。
优选的，所述心肌肌钙蛋白I抗体(cTnl)小部分(约3％)游离于包浆中，为可溶性，所述心肌肌钙蛋白I抗体(cTnl)大部分以结构蛋白的形式固定在肌原纤维上，为不溶性，所述心肌肌钙蛋白I抗体(cTnl)在心肌细胞膜完整的状态下，不能透过细胞膜进入血循环，血液中心肌肌钙蛋白I抗体(cTnl)的含量很低，当细胞膜的完整性因缺血或缺氧等受到破坏时，游离的心肌肌钙蛋白I抗体(cTnl)可迅速透过细胞膜进入血流，而结合的心肌肌钙蛋白I抗体(cTnl)则逐渐分解出来，成为游离的心肌肌钙蛋白I抗体(cTnl)。
优选的，所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量＜0.1ng/ml。
优选的，所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量在0.1-0.5ng/ml。
优选的，所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量在0.5-25ng/ml。
优选的，所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量＞25ng/ml。
与现有技术相比，本发明的有益效果是：
本发明中，心肌肌钙蛋白I抗体(cTnl)抗体采用体外纯化细胞培养上清得到，与腹水来源的心肌肌钙蛋白I抗体(cTnl)相比，细胞体外培养生产的抗体避免了腹水中其它鼠源成分的干扰，产品质量更好，同时批次间差异更小，更稳定。
附图说明
图1为本发明免疫荧光平台样本符合率(用心肌肌钙蛋白I抗体(cTnl)抗体制备心肌肌钙蛋白I抗体(cTnl)免疫荧光纸条，检测雅培赋值样本30例(浓度范围0.05-45ng/ml)，样本符合率R2>98％)；
图2是将心肌肌钙蛋白l稀释成5ng/ml-20ng/ml和0.1ng/ml-2ng/ml两个浓度水平的样本，对每个样品重复测定3次，根据公式B＝(M-T)/Tx100％(式中B为相对偏差,M为测量浓度的均值，T为标定浓度)计算相对偏差,相对偏差应不大于15％；
图3是用心肌肌钙蛋白l浓度为0.1ng/ml-2ng/ml和5ng/ml-20ng/ml的两个水平的质控品为样本，重复测定各10次,计算测量值的平均值和标准差,按公式CV＝(标准差/平均值)x100％，计算变异系数(CV),结果应不大于15％。
具体实施方式
实施例1：本发明提供一种技术方案：
一种高特异性的心肌肌钙蛋白I抗体及其制备方法和用途，心肌肌钙蛋白I抗体包括210个氨基酸组成，分子量23.9，等电点9.87，是一种富含α螺旋的蛋白质，有5个螺旋区域，呈伸展式构型，心肌肌钙蛋白I抗体(cTnl)为鼠单抗，体外纯化细胞培养上清获得，用于免疫发光和化学发光。
所述心肌肌钙蛋白I抗体(cTnl)小部分(约3％)游离于包浆中，为可溶性，所述心肌肌钙蛋白I抗体(cTnl)大部分以结构蛋白的形式固定在肌原纤维上，为不溶性，所述心肌肌钙蛋白I抗体(cTnl)在心肌细胞膜完整的状态下，不能透过细胞膜进入血循环，血液中心肌肌钙蛋白I抗体(cTnl)的含量很低，当细胞膜的完整性因缺血或缺氧等受到破坏时，游离的心肌肌钙蛋白I抗体(cTnl)可迅速透过细胞膜进入血流，而结合的心肌肌钙蛋白I抗体(cTnl)则逐渐分解出来，成为游离的心肌肌钙蛋白I抗体(cTnl)，可以实际的使用过程中通过血液中心肌肌钙蛋白I抗体(cTnl)的含量对疾病进行检测；所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量＜0.1ng/ml，可以排除心肌损伤的问题；所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量在0.1-0.5ng/ml，心肌损伤低风险，需结合多种诊断结果及临床症状综合判断；所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量在0.5-25ng/ml，心肌损伤高风险；所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量＞25ng/ml，严重心肌损伤。
心肌肌钙蛋白I抗体(cTnl)为心肌组织特有的一种调节蛋白，可抑制肌球蛋白与肌动蛋白结合，在心肌收缩过程中起重要作用，在急性心肌梗死(acute myocardialinfaction,AMI)早期心肌肌钙蛋白I抗体(cTnl)水平变化幅度较大,具有较高的敏感性，且心肌肌钙蛋白I抗体(cTnl)不在任何类型的骨骼肌中表达，具有较高的心肌细胞特异性。正常人血清中心肌肌钙蛋白I抗体(cTnl)平均浓度<0.06ng/ml，当心肌轻微损伤时其浓度明显升高，可达100-1300ng/ml，心肌肌钙蛋白I抗体(cTnl)诊断心肌梗塞的特异性为96％，敏感性为97％，因此，心肌肌钙蛋白I抗体(cTnl)是心肌细胞损伤最敏感和最特异性的血清标志物之一,加之心肌肌钙蛋白I抗体(cTnl)具有诊断阈值明确、窗口期宽以及检测快速等优点，已逐渐判断AMI患者心肌细胞损伤的主要生化指标。
实施例2：本发明提供一种技术方案：
一种高特异性的心肌肌钙蛋白I抗体及其制备方法和用途，心肌肌钙蛋白I抗体包括210个氨基酸组成，分子量23.9，等电点9.87，是一种富含α螺旋的蛋白质，有5个螺旋区域，呈伸展式构型，心肌肌钙蛋白I抗体(cTnl)为鼠单抗，体外纯化细胞培养上清获得，用于免疫发光和化学发光。
所述心肌肌钙蛋白I抗体(cTnl)小部分(约3％)游离于包浆中，为可溶性，所述心肌肌钙蛋白I抗体(cTnl)大部分以结构蛋白的形式固定在肌原纤维上，为不溶性，所述心肌肌钙蛋白I抗体(cTnl)在心肌细胞膜完整的状态下，不能透过细胞膜进入血循环，血液中心肌肌钙蛋白I抗体(cTnl)的含量很低，当细胞膜的完整性因缺血或缺氧等受到破坏时，游离的心肌肌钙蛋白I抗体(cTnl)可迅速透过细胞膜进入血流，而结合的心肌肌钙蛋白I抗体(cTnl)则逐渐分解出来，成为游离的心肌肌钙蛋白I抗体(cTnl)，可以实际的使用过程中通过血液中心肌肌钙蛋白I抗体(cTnl)的含量对疾病进行检测；所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量＜0.1ng/ml，恢复较好；所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量在0.1-0.5ng/ml，心肌损伤低风险，恢复良好需要复查；所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量在0.5-25ng/ml，恢复效果一般；所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量＞25ng/ml，恢复效果十分不理想。
静脉注入溶栓药物是近年来常用的急性心肌梗死治疗方法，在治疗后判断是否出现冠状脉血管再灌通，梗塞开通后，血流进入病变部位,将游离的心肌肌钙蛋白I抗体(cTnl)或心肌肌钙蛋白I抗体(cTnl)冲洗入血液而出现第一个峰，此后可观察到第二个较小的峰，检测心肌肌钙蛋白I抗体(cTnl)可以对这类溶栓药物的作用进行判断。
本文中应用了具体个例对本发明的原理及实施方式进行了阐述，以上实例的说明只是用于帮助理解本发明的方法及其核心思想。以上所述仅是本发明的优选实施方式，应当指出，由于文字表达的有限性，而客观上存在无限的具体结构，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进、润饰或变化，也可以将上述技术特征以适当的方式进行组合；这些改进润饰、变化或组合，或未经改进将发明的构思和技术方案直接应用于其它场合的，均应视为本发明的保护范围。
Claims (6)
1.一种高特异性的心肌肌钙蛋白I抗体及其制备方法，其特征在于：心肌肌钙蛋白I抗体包括210个氨基酸组成，分子量23.9，等电点9.87，是一种富含α螺旋的蛋白质，有5个螺旋区域，呈伸展式构型，心肌肌钙蛋白I抗体(cTnl)为鼠单抗，体外纯化细胞培养上清获得，用于免疫发光和化学发光。
2.根据权利要求1所述的一种高特异性的心肌肌钙蛋白I抗体用途，其特征在于：所述心肌肌钙蛋白I抗体(cTnl)小部分(约3％)游离于包浆中，为可溶性，所述心肌肌钙蛋白I抗体(cTnl)大部分以结构蛋白的形式固定在肌原纤维上，为不溶性，所述心肌肌钙蛋白I抗体(cTnl)在心肌细胞膜完整的状态下，不能透过细胞膜进入血循环，血液中心肌肌钙蛋白I抗体(cTnl)的含量很低，当细胞膜的完整性因缺血或缺氧等受到破坏时，游离的心肌肌钙蛋白I抗体(cTnl)可迅速透过细胞膜进入血流，而结合的心肌肌钙蛋白I抗体(cTnl)则逐渐分解出来，成为游离的心肌肌钙蛋白I抗体(cTnl)。
3.根据权利要求2所述的一种高特异性的心肌肌钙蛋白I抗体用途，其特征在于：所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量＜0.1ng/ml。
4.根据权利要求2所述的一种高特异性的心肌肌钙蛋白I抗体用途，其特征在于：所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量在0.1-0.5ng/ml。
5.根据权利要求2所述的一种高特异性的心肌肌钙蛋白I抗体用途，其特征在于：所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量在0.5-25ng/ml。
6.根据权利要求2所述的一种高特异性的心肌肌钙蛋白I抗体用途，其特征在于：所述心肌肌钙蛋白I抗体(cTnl)在血液中的含量＞25ng/ml。
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