提出了一种基于纳米金标记的阳极溶出伏安免疫分析方法。免疫反应在聚苯乙烯微孔板中以夹心分析模式进行,通过物理吸附将兔抗人免疫球蛋白 A(IgA)抗体固定于微孔板上,与相应抗原 IgA 发生免疫反应后,再通过夹心模式捕获相应的纳米金标...提出了一种基于纳米金标记的阳极溶出伏安免疫分析方法。免疫反应在聚苯乙烯微孔板中以夹心分析模式进行,通过物理吸附将兔抗人免疫球蛋白 A(IgA)抗体固定于微孔板上,与相应抗原 IgA 发生免疫反应后,再通过夹心模式捕获相应的纳米金标记的羊抗人 IgA 抗体,然后再与金标羊抗人 IgA 抗体和金标兔抗羊二抗形成的免疫复合物反应,在微孔板上引入大量的纳米金,将金溶解后,在碳糊电极上用阳极溶出伏安法(ASV)对金离子进行检测,溶出峰电流的大小间接与待分析物 IgA 的浓度成正比。对免疫分析的一些实验条件进行了优化。阳极溶出峰电流与 IgA 的浓度在0.018 mg/L～181 mg/L 范围内呈良好的线性关系,检测下限为10μg/L。将该分析方法应用于人血清中 IgA 浓度的测定,取得了满意结果。展开更多
A nanoparticle-based potentiometric immunoassay was designed for the sensitive detection of squamous cell carcinoma antigen(SCCA;cervical carcinoma marker) on a portable pH meter coupling enzyme-labeled hybridization ...A nanoparticle-based potentiometric immunoassay was designed for the sensitive detection of squamous cell carcinoma antigen(SCCA;cervical carcinoma marker) on a portable pH meter coupling enzyme-labeled hybridization chain reaction(HCR) with two alternating hairpin DNA probes for the signal amplification. Initially, a sandwich-type immunoreaction was carried out between anti-SCCA capture antibody-conjugated magnetic bead and detection antibody/initiator strand-coated gold nanoparticle(AuNP). Then, the HCR reaction was readily executed between two glucose oxidase(GOx)-labeled hairpins through the initiator strand to form numerous GOx concatamers on the AuNP via the long nicked double-helix. The concatenated GOx oxidized glucose into gluconic acid and hydrogen peroxide, thus resulting in the pH change of the detection solution on a handheld pH meter. Several labeling protocols including GOx-antibody, GOx-AuNP-antibody and GOx-HCR-AuNP-antibody were investigated for detection of target SCCA, and improved analytical features were obtained with the immune-HCR assay. Under optimum conditions, the immune-HCR assay exhibited good pH responses for the determination of SCCA at a concentration as low as 5.7 pg/mL. Additionally, the immune-HCR assay had good precision and reproducibility, high specificity, and acceptable accuracy for analyzing human serum specimens.展开更多
文摘提出了一种基于纳米金标记的阳极溶出伏安免疫分析方法。免疫反应在聚苯乙烯微孔板中以夹心分析模式进行,通过物理吸附将兔抗人免疫球蛋白 A(IgA)抗体固定于微孔板上,与相应抗原 IgA 发生免疫反应后,再通过夹心模式捕获相应的纳米金标记的羊抗人 IgA 抗体,然后再与金标羊抗人 IgA 抗体和金标兔抗羊二抗形成的免疫复合物反应,在微孔板上引入大量的纳米金,将金溶解后,在碳糊电极上用阳极溶出伏安法(ASV)对金离子进行检测,溶出峰电流的大小间接与待分析物 IgA 的浓度成正比。对免疫分析的一些实验条件进行了优化。阳极溶出峰电流与 IgA 的浓度在0.018 mg/L～181 mg/L 范围内呈良好的线性关系,检测下限为10μg/L。将该分析方法应用于人血清中 IgA 浓度的测定,取得了满意结果。
基金financial support from the National Natural Science Foundation of China (No. 21675029)the Health-Education Joint Research Project of Fujian Province (No. WKJ2016-2-15)
文摘A nanoparticle-based potentiometric immunoassay was designed for the sensitive detection of squamous cell carcinoma antigen(SCCA;cervical carcinoma marker) on a portable pH meter coupling enzyme-labeled hybridization chain reaction(HCR) with two alternating hairpin DNA probes for the signal amplification. Initially, a sandwich-type immunoreaction was carried out between anti-SCCA capture antibody-conjugated magnetic bead and detection antibody/initiator strand-coated gold nanoparticle(AuNP). Then, the HCR reaction was readily executed between two glucose oxidase(GOx)-labeled hairpins through the initiator strand to form numerous GOx concatamers on the AuNP via the long nicked double-helix. The concatenated GOx oxidized glucose into gluconic acid and hydrogen peroxide, thus resulting in the pH change of the detection solution on a handheld pH meter. Several labeling protocols including GOx-antibody, GOx-AuNP-antibody and GOx-HCR-AuNP-antibody were investigated for detection of target SCCA, and improved analytical features were obtained with the immune-HCR assay. Under optimum conditions, the immune-HCR assay exhibited good pH responses for the determination of SCCA at a concentration as low as 5.7 pg/mL. Additionally, the immune-HCR assay had good precision and reproducibility, high specificity, and acceptable accuracy for analyzing human serum specimens.
