多肽在金纳米粒子表面偶联反应效率研究

通过1-(3-二甲氨基丙基)-3-乙基碳二亚胺(EDC)与N-羟基琥珀酰亚胺(NHS)活化反应将多肽偶联到金纳米粒子表面,采用荧光光谱研究其形成的酰胺键的反应偶联效率.考察了实验条件,包括缓冲液的种类(HEPES、Tris-HCl、硼酸、PBS缓冲液)、pH值(pH 6.5~9.0)、缓冲液浓度(10, 25, 40和50 mmol/L)、NHS和EDC的浓度(NHS 0.2~1.0 mol/L,EDC 0.01~0.5 mol/L)及二者比例(0,0.5,1.0,2.0和2.5)、偶联反应时间(4, 8, 12, 24和36 h)等对偶联效率的影响,筛选出最佳实验条件.实验结果表明, 25 mmol/L 4-羟乙基哌嗪乙磺酸(HEPES)缓冲液(pH 7.0), NHS/EDC浓度为0.4 mol/L/0.2 mol/L和24 h的反应时间为EDC-NHS活化反应将多肽偶联到金纳米粒子上的最佳实验条件.本研究结果可为相关研究提供技术参考.

Study of Efficiency of Coupling Peptides with Gold Nanoparticles

Fluorescence spectroscopy was used to investigate the efficiency of coupling peptides to gold nanoparticles via 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-N-hydroxysuccinimide (EDC-NHS).The experiment conditions including buffer solution, pH value and concentrations of buffer solution, concentrations of NHS and EDC, concentration ratios of NHS to EDC, and coupling reaction time on the coupling efficiency were investigated and optimized.The experimental results indicated that the optimized experimental conditions were as follows: 25 mmol/L HEPES buffer solution, pH 7.0;2∶1 of concentration ratio of NHS to EDC, 0.4 mol/L NHS, 0.2 mol/L EDC, and coupling reaction time of 24 h.This study may provide references for the relative research involving coupling peptide or protein with gold nanoparticles