We describe for the first time the synthesis and the optimal conditions for protein labeling with a new fluorescent probe,5-chlorosulfoyl-2-thenoyltrifluoroacetone(CTTA),whicb forms a highly fluorescent conplex with E...We describe for the first time the synthesis and the optimal conditions for protein labeling with a new fluorescent probe,5-chlorosulfoyl-2-thenoyltrifluoroacetone(CTTA),whicb forms a highly fluorescent conplex with Eu^(3+) when conjugated to protein.The labeled proteins were characterized by absorbance and fluorescence measurements and the effect of labeling on the biological activity of sone proteins was also studied.It is shown that the new label is suitable for applications in time-resolved fluoroimmunoassay.展开更多
A series of Eu( Ⅲ )/Y( Ⅲ ) complexes of terephthalic acid(TPA) with 2-thenoyltrifluoroacetone(HTTA) and trioctylphosphine oxide(TPPO) were synthesized. Compositions of these complexes are revealed to be Eu2(1-x)Y2x(...A series of Eu( Ⅲ )/Y( Ⅲ ) complexes of terephthalic acid(TPA) with 2-thenoyltrifluoroacetone(HTTA) and trioctylphosphine oxide(TPPO) were synthesized. Compositions of these complexes are revealed to be Eu2(1-x)Y2x(TPA)(TTA)4- (TPPO)4 or Eu1-xYx(TPA)(TTA)(TPPO)2. Their IR spectra, fluorescent spectra and the thermal and optical stability were studied. The fluorescent excitation spectra of these complexes show more broad excitation bands than those of Eu2(TPA)3(TPPO)4 and Eu(TTA)3(TPPO)2 corresponding to their formations. In addition, both the binuclear complex EuY(TPA)(TTA)4(TPPO)4 and the chain complex Eu0.4Y0.6(TPA)(TTA)(TPPO)2 present higher thermal stability and better optical stability than the mononuclear complex Eu(TTA)3(TPPO)2 does. And their thermal and optical stabilities are preferably interpreted from the binuclear structure together with the chain polynuclear structure of the complexes. The fluorescence enhancement of cofluorescence Y3+ ions to the Eu(Ⅲ) complexes is clear. The optimum content of Y3+ is 0.6 (molar fraction) for the chain complexes Eu1-xYx(TPA)(TTA)- (TPPO)2 and 0.5 for the binuclear complexes Eu2(1-x)Y2x(TPA)(TTA)4(TPPO)4. The formation of polynuclear structure of the complex Eu1-xYx(TPA)(TTA)(TPPO)2 appears to be responsible for the good cofluorescence effect of Y3+ ions.展开更多
芝麻是八大类食物过敏原之一,快速准确识别芝麻过敏原对预防其过敏有重要意义。核酸适配体可以高效识别靶标过敏原,在过敏原检测中有良好的应用前景。为了获得芝麻主要过敏原Ses i 2的特异性核酸适体,本研究以Ses i 2为靶标,通过磁珠筛...芝麻是八大类食物过敏原之一,快速准确识别芝麻过敏原对预防其过敏有重要意义。核酸适配体可以高效识别靶标过敏原,在过敏原检测中有良好的应用前景。为了获得芝麻主要过敏原Ses i 2的特异性核酸适体,本研究以Ses i 2为靶标,通过磁珠筛选法(磁珠-SELEX)开展10轮筛选,经由高通量测序获得6条候补序列(S1~S6),并进行家族性、同源性分析及二级结构预测。结果表明,6条候选核酸适体的重复率可达46.38%,其自由能在-9.02到-2.47 kcal·moL^(-1)之间,根据自由能能量稳定原则,S1和S5吉布斯自由能最低最稳定,分别为-6.70和-9.02 kcal·moL^(-1)。利用ELISA试验进行亲和力测试,结果表明核酸适体S1和S2的亲和能力较强,S1:KD=67.02 nmol·L^(-1),R2=0.925 8,S2:KD=97.65 nmol·L^(-1),R2=0.795 1。核酸适体S1与过敏原Ses i 2的结合力和其他过敏原蛋白相比有显著差异,可视为具有特异性。本研究最终获得一条兼具良好亲和力和特异性的核酸适体S1,为芝麻过敏原快速检测提供了技术支撑。展开更多
基金Supported by National Natural Science Foundation of China.
文摘We describe for the first time the synthesis and the optimal conditions for protein labeling with a new fluorescent probe,5-chlorosulfoyl-2-thenoyltrifluoroacetone(CTTA),whicb forms a highly fluorescent conplex with Eu^(3+) when conjugated to protein.The labeled proteins were characterized by absorbance and fluorescence measurements and the effect of labeling on the biological activity of sone proteins was also studied.It is shown that the new label is suitable for applications in time-resolved fluoroimmunoassay.
基金Project(20576142) supported by the National Natural Science Foundation of ChinaProject(05B075) supported by the Foundation of Hunan Education Bureau for Young Scholars, China
文摘A series of Eu( Ⅲ )/Y( Ⅲ ) complexes of terephthalic acid(TPA) with 2-thenoyltrifluoroacetone(HTTA) and trioctylphosphine oxide(TPPO) were synthesized. Compositions of these complexes are revealed to be Eu2(1-x)Y2x(TPA)(TTA)4- (TPPO)4 or Eu1-xYx(TPA)(TTA)(TPPO)2. Their IR spectra, fluorescent spectra and the thermal and optical stability were studied. The fluorescent excitation spectra of these complexes show more broad excitation bands than those of Eu2(TPA)3(TPPO)4 and Eu(TTA)3(TPPO)2 corresponding to their formations. In addition, both the binuclear complex EuY(TPA)(TTA)4(TPPO)4 and the chain complex Eu0.4Y0.6(TPA)(TTA)(TPPO)2 present higher thermal stability and better optical stability than the mononuclear complex Eu(TTA)3(TPPO)2 does. And their thermal and optical stabilities are preferably interpreted from the binuclear structure together with the chain polynuclear structure of the complexes. The fluorescence enhancement of cofluorescence Y3+ ions to the Eu(Ⅲ) complexes is clear. The optimum content of Y3+ is 0.6 (molar fraction) for the chain complexes Eu1-xYx(TPA)(TTA)- (TPPO)2 and 0.5 for the binuclear complexes Eu2(1-x)Y2x(TPA)(TTA)4(TPPO)4. The formation of polynuclear structure of the complex Eu1-xYx(TPA)(TTA)(TPPO)2 appears to be responsible for the good cofluorescence effect of Y3+ ions.
文摘芝麻是八大类食物过敏原之一,快速准确识别芝麻过敏原对预防其过敏有重要意义。核酸适配体可以高效识别靶标过敏原,在过敏原检测中有良好的应用前景。为了获得芝麻主要过敏原Ses i 2的特异性核酸适体,本研究以Ses i 2为靶标,通过磁珠筛选法(磁珠-SELEX)开展10轮筛选,经由高通量测序获得6条候补序列(S1~S6),并进行家族性、同源性分析及二级结构预测。结果表明,6条候选核酸适体的重复率可达46.38%,其自由能在-9.02到-2.47 kcal·moL^(-1)之间,根据自由能能量稳定原则,S1和S5吉布斯自由能最低最稳定,分别为-6.70和-9.02 kcal·moL^(-1)。利用ELISA试验进行亲和力测试,结果表明核酸适体S1和S2的亲和能力较强,S1:KD=67.02 nmol·L^(-1),R2=0.925 8,S2:KD=97.65 nmol·L^(-1),R2=0.795 1。核酸适体S1与过敏原Ses i 2的结合力和其他过敏原蛋白相比有显著差异,可视为具有特异性。本研究最终获得一条兼具良好亲和力和特异性的核酸适体S1,为芝麻过敏原快速检测提供了技术支撑。
