超高容量氧化石墨烯/碳纳米管气凝胶用于血红蛋白的吸附分离

doi:10.7503/cjcu20150175

摘要/Abstract

摘要：

采用冷冻干燥技术制备了三维氧化石墨烯/碳纳米管(GO/CNTs)气凝胶, 在疏水作用下, 该气凝胶对蛋白质表现出良好的吸附性能. 在pH=8.0时, GO/CNTs气凝胶对血红蛋白的吸附效率很高, 而对白蛋白基本不吸附, 从而可实现血红蛋白的选择性吸附分离. 在最佳吸附条件下, GO/CNTs气凝胶对血红蛋白的吸附容量高达3793.3 mg/g, 是去除高丰度血红蛋白的高效材料. 吸附后的血红蛋白可以使用pH=11.5的Britton-Robisin缓冲液有效洗脱, 回收率为67%. 圆二色光谱结果表明, GO/CNTs气凝胶具有良好的生物相容性, 其表面发生的吸附/洗脱过程对血红蛋白的构象基本无影响. 将GO/CNTs气凝胶用于人全血样品中血红蛋白的选择性分离, SDS-聚丙烯酰胺凝胶电泳结果表明得到了较高纯度的血红蛋白.

关键词: 氧化石墨烯, 碳纳米管, 气凝胶, 血红蛋白, 分离

Abstract:

3-Dimensional graphene oxide/carbon nanotubes(GO/CNTs) aerogel was prepared via freeze drying technique, and the as-prepared GO/CNTs aerogel exhibits excellent adsorption performance towards protein species under hydrophobic interaction. At pH=8.0, high adsorption efficiency of hemoglobin(Hb) was achieved, while virtually no adsorption of serum album was observed. This offers the possibility to realize the selective isolation of Hb from complex matrices. An adsorption capacity up to 3793.3 mg/g was achieved for the GO/CNTs aerogel, suggesting the great potential of this aerogel in the removal of abundant Hb. The adsorbed Hb could be readily stripped using Britton-Robinson buffer of pH=11.5, giving rise to a recovery of 67%. Circular dichroism spectra indicated that the adsorption/desorption on the surface of GO/CNTs aerogel poses virtually no effect on the configuration of Hb. The GO/CNTs aerogel is practically applied to the selective isolation of Hb from human blood and SDS-PAGE assays demonstrate high purity of hemoglobin in the eluate.

Key words: Graphene oxide, Carbon nanotubes, Aerogel, Hemoglobin, Isoaltion

中图分类号:

O652.6

TrendMD:

陈旭伟, 郑旭, 王晓峰, 王建华. 超高容量氧化石墨烯/碳纳米管气凝胶用于血红蛋白的吸附分离. 高等学校化学学报, 2015, 36(8): 1498.

CHEN Xuwei, ZHENG Xu, WANG Xiaofeng, WANG Jianhua. Graphene/Carbon Nanaotube Aerogel with Ultra-high Adsorption Capacity for the Isolation of Hemoglobin^†. Chem. J. Chinese Universities, 2015, 36(8): 1498.

图/表 9

Fig.1 Schematic diagram for the preparation of GO/CNTs aerogel

Fig.2 SEM(A—C) and TEM(D—F) images of GO(A, D), CNTs(B, E) and GO/CNTs aerogel(C, F)

Fig.3 pH-dependent adsorption of protein species onto the GO/CNTs aerogela. Hb; b. BSA. Protein: 100 mg/L, 1.0 mL;GO/CNTs aerogel: 0.1 mg; adsorption time: 25 min.

Fig.4 Effect of ionic strength on the adsorption efficiencyHb: 100 mg/L, 1.0 mL; GO/CNTs aerogel: 0.1 mg;pH=8.0; adsorption time: 25 min.

Fig.5 Adsorption isotherms of Hb by GO/CNTs aerogel at pH=6.0(a) and pH=8.0(b)GO/CNTs aerogel: 0.1 mg; adsorption time: 25 min.

Table 1 Performance of various materials on the adsorption of Hb

Solid material	Adsorption time/min	Adsorption capacity/(mg·g^-1)	Reference
BHb-imprinted magnetic nanoparticles	30	4.65	[27]
Molecularly imprinted chitosan beads	1020	32.5	[28]
Organic-inorganic hybrid silica	300	43.6	[29]
Magnetic molecularly imprinted nanoparticles	180	77.6	[30]
Imidazolium-modified polystyrene	10	23.6	[31]
Imidazolium-modified sulfonated polyetheretherketone	10	31.6	[32]
PPimPF₆/TiO₂ nanocomposites	20	122.3	[33]
La³⁺ modified zeolite	30	31.8	[34]
Keggin-type phosphomolybdate	20	55.86	[35]
GO/SiO₂ composites	30	50.5	[23]
La-MOF-GO composite	30	193	[36]
GO/CNTs aerogel	25	3793.3^*	This work

Fig.6 Elution efficiency of hemoglobin using various stripping reagents(A) and the effect of B-R buffer pH on the elution efficiency(B)(A) a. Glycine-NaOH(0.01 mol/L, pH=9.4); b. Borax-NaOH(0.01 mol/L, pH=10.1); c. B-R(0.01 mol/L, pH=10.0); d. Na3PO4(0.01 mol/L); e. Tris-HCl(0.01 mol/L, pH=8.9). Hb: 100 mg/L; adsorption time: 25 min; elution time: 30 min.

Fig.7 CD spectra of Hb in B-R buffer---- Eluate; ——Hb standard.

Fig.8 SDS-PAGE assay resultsLane 1: molecular weight standards; lane 2: 500-fold diluted human whole blood without pretreatment; lane 3: 500-fold diluted human whole blood after adsorption by the GO/CNTs aerogel; lane 4: Hb recovered from the GO/CNTs aerogel surface; lane 5: Hb standard solution of 64 mg/L.

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