Quantification of human embryonic ζ-globin chains in Southeast Asian deletion (--SEA) carriers
Quantification of human embryonic ζ-globin chains in Southeast Asian deletion (--SEA) carriers
复制标题
东南亚缺失 (--SEA) 携带者中人类胚胎 γ 珠蛋白链的定量
DOI：
10.1136/jcp-2022-208159
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发表时间：
2022
影响因子：
3.4
通讯作者：
Xia
中科院分区：
文献类型：
--
作者：
Yuhua Ye;Guoying Sun;Zhe Ren;Yidan Liang;Hualei Luo;Peng Lin;Xingmin Wang;Zejun Dong;Li Huang;Lang Qin;Wenfang Yu;Ge Wang;Yuqiu Zhou;Jia Tang;Jiwu Lou;Yanhui Liu;Xianqi Zeng;Yajun Chen;Yihong Li;Qianqian Zhang;Jin Huang;Ping Zhu;Liang Lin;Xinhua Zhang;Xia
目的 胚胎 δ-珠蛋白的再激活是 α-地中海贫血基因治疗的一种有前途的策略。然而，作为 α-地中海贫血携带者和患者的数量性状的 δ-球蛋白的定量仍不完全清楚。在本研究中，我们旨在建立一种可靠的方法来定量α-地中海贫血携带者中的δ-珠蛋白，然后进行群体研究以研究其表达模式。方法从Hb Bart胎儿的脐带血溶血液中纯化δ-珠蛋白作为单体，然后进行绝对蛋白质定量，然后通过内部ELISA系统进行测试并引入作为蛋白质标准品。然后用于对 6179 名个体的外周血样本进行大规模定量。最后，通过测量第二组 141-SEA/αα 携带者中的 z-珠蛋白表达，引入液相色谱-串联质谱 (LC-MS/MS) 作为独立的验证方法。结果证明 ELISA 系统在区分具有不同程度的 z-珠蛋白的个体方面很灵敏。对这一--SEA/αα 携带者群体的大规模定量研究表明，δ-珠蛋白表达具有高度多样性，范围为 0.00155 g/L 至 1.48778 g/L。 ELISA 和 LC-MS/MS 之间显着正相关（R=0.400，p结论）我们的研究报告了可靠的 δ-珠蛋白定量方法，并呈现了--SEA/αα 携带者群体中 δ-珠蛋白的表达模式，这可能为后续 α-地中海贫血延迟性血红蛋白转换机制的基因型-表型研究奠定基础。
AimsReactivation of embryonic ζ-globin is a promising strategy for genetic treatment of α-thalassaemia. However, quantification of ζ-globin as a quantitative trait in α-thalassaemia carriers and patients remains incompletely understood. In this study, we aimed to set up a reliable approach for the quantification of ζ-globin in α-thalassaemia carriers, followed by a population study to investigate its expression patterns.Methodsζ-globin was purified as monomers from cord blood haemolysate of a Hb Bart’s fetus, followed by absolute protein quantification, which was then tested by in-house ELISA system and introduced as protein standard. It was then used for large-scale quantification in peripheral blood samples from 6179 individuals. Finally, liquid chromatography-tandem mass spectrometry (LC-MS/MS) introduced as an independent validating approach by measuring ζ-globin expression in a second cohort of 141-SEA/αα carriers.ResultsThe ELISA system was proved sensitive in distinguishing individuals with varied extent of ζ-globin. Large scale quantitative study of this --SEA/αα carrier cohort indicated the high diversity of ζ-globin expression ranging from 0.00155 g/L to 1.48778 g/L. Significant positive correlation between ELISA and LC-MS/MS (R=0.400, pConclusionOur study has reported reliable approaches for the quantification of ζ-globin and presented the expression patterns of ζ-globin among the --SEA/αα carrier population, which might lay a foundation on subsequent genotype–phenotype studies on mechanisms of delayed haemoglobin switch in α-thalassaemia.