用生长因子包被颗粒和流式细胞术研究PDGF-BB适体结合

Q3 Biochemistry, Genetics and Molecular Biology Journal of Advanced Biotechnology and Experimental Therapeutics Pub Date : 2023-01-01 DOI:10.5455/jabet.2023.d122
Supavinee Sukcharoen, Paphada Watcharapo, Kanokkorn Vichitsakul, Jiraporn Arunpanichlert, P. Rotkrua, B. Soontornworajit
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引用次数: 0

摘要

血小板衍生生长因子bb (platelet derived growth factor- bb, PDGF-BB)是一种由血小板、内皮细胞和癌细胞分泌的多功能生长因子,是一种高效、准确的生物标志物。PDGF- BB已用于各种治疗应用。通过特定的结合配体调节其生物活性可以增强PDGF-BB的功能。适配体是一种单链寡核苷酸,可选择性结合PDGF-BB,是我们感兴趣的分子。本研究旨在评价生长因子包被颗粒与PDGF-BB适配体的结合。ELISA检测pdgf - bb包被颗粒,流式细胞术检测适配体结合评价。为优化结合评价条件,采用流式细胞仪对PDGF-BB、聚苯乙烯颗粒和适配体的洗涤周期和浓度进行了研究。流式细胞术结果显示,PDGF-BB的浓度在荧光颗粒数上呈现最佳水平。随着洗涤周期的缩短和颗粒浓度的降低,荧光颗粒的百分比增加。荧光粒子的百分比也随适配体浓度的增加而增加。利用流式细胞仪检测生长因子包被颗粒的荧光信号是研究生物分子结合的一种很有前途的方法。
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Investigation of PDGF-BB aptamer binding using growth factor-coated particles and flow cytometry
Platelet-derived growth factor-BB (PDGF-BB), a multifunctional growth factor secreted by platelets, endothelial and cancer cells, functions as an efficient and accurate biomarker. PDGF- BB has been employed in various therapeutic applications. Tuning its bioactivity via a specific binding ligand could enhance the PDGF-BB functionality. Aptamer, a single-stranded oligonucleotide that exhibits a selective binding to PDGF-BB, is the molecule of our interest. This research aims to evaluate the PDGF-BB aptamer binding by growth factor-coated particles. PDGF-BB-coated particles were confirmed by ELISA, and aptamer binding evaluation was investigated by flow cytometry. To optimize the condition for binding evaluation, the washing cycle and concentration of PDGF-BB, polystyrene particle and aptamer were studied using flow cytometer. Based on the flow cytometry results, the concentration of PDGF-BB showed an optimal level on the number of fluorescent particles. The percentage of fluorescent particles increased when the washing cycle was reduced and the particle concentration was decreased. The percentage of fluorescent particles also increased proportionally with the aptamer concentration. Using a flow cytometer to detect fluorescent signal from growth factor-coated particle is a promising strategy to study biomolecular binding.
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来源期刊
Journal of Advanced Biotechnology and Experimental Therapeutics
Journal of Advanced Biotechnology and Experimental Therapeutics Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
1.90
自引率
0.00%
发文量
41
审稿时长
8 weeks
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