使用抗体衍生标签进行蛋白质表达的流细胞计分析,其次是CITE-Seq
Xiaoshan Shi1, Wei Fan2, Majid Mehrpouyan2
1Applied Research & Technology, Medical Scientific Affairs, BD Biosciences, San Jose, California, USA.
概括
这项研究引入了一种新型的流动细胞计量试验,用于验证通过测序 (CITE-Seq) 进行转录体和表皮细胞的细胞索引的抗体衍生标签 (ADT). 该方法提高了ADT标记物选择和细胞分类效率,用于多组学分析.
科学领域:
- 单细胞多组的多组学
- 免疫型定型 免疫型定型
- 分子生物学分子生物学
背景情况:
- 通过测序对转录组和表皮细胞进行细胞索引 (CITE-Seq) 是一种流行的单细胞表型化技术.
- 目前的CITE-Seq协议缺乏详细的方法来选择抗体衍生标签 (ADT) 标记物并确保试剂的性能.
- 这种差距阻碍了对细胞异质性和动态的准确研究.
研究的目的:
- 为CITE-Seq.开发一种新的,用户友好的多重流量代理测试.
- 为了使多个蛋白质标记物的同时测量使用ADT试剂和染料-oligo结合物.
- 在测序之前指导ADT标记物选择并确认蛋白质表达.
主要方法:
- 使用ADT试剂和补充染料-基合物开发了一种多重流细胞计测定.
- 该试验允许对蛋白质标记物表达的特定结合和多重评估.
- 标记的细胞使用基于下游多组学的染料-藻光学的流动细胞计分类.
主要成果:
- 该试验有效地以多重方式评估蛋白质标记物表达.
- 它可以有效地分类特定细胞群,如自然杀手细胞和T细胞.
- 这种方法避免了与传统的共同染色相关的潜在标记物分辨率问题.
结论:
- 开发的工作流简化了ADT标记物选择,并提高了CITE-Seq.的细胞分类效率.
- 这种方法为提高单细胞多组学研究的质量和准确性提供了一个可行的选择.
- 它促进了强大的细胞表型和转录基因分析.
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