Antibody fluorescein-doped silica nanobioconjugates for the ultrasensitive detection of prostate-specific antigen†

IF 3.5 Q2 CHEMISTRY, ANALYTICAL
Tumelo Msutu, Omotayo Adeniyi and Philani Mashazi
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Abstract

Herein, we report the bioconjugation of anti-prostate-specific antigen polyclonal antibodies (pAb) onto the fluorescein-doped silica nanoparticles to detect prostate-specific antigen (PSA). Fluorescein-isothiocyanate (FITC), a fluorescent dye, was reacted with (3-aminopropyl)triethoxysilane (APTES) to form the FITC-APTES organosilane precursor. FITC-APTES was mixed with tetraethoxysilane (TEOS) to form fluorescent silica nanoparticles (FITC@SiO2NPs) containing 3% and 6% of dye loading. The silica nanoparticles prevented the dye from leaching and promoted the fluorescent signal amplification for the detection of PSA. Phenylboronic acid (PBA) was coated onto the fluorescent silica nanoparticles for the oriented antibody immobilization via the boronate ester to form FITC@SiO2-PBA-pAb. The fluorescent silica nanobioconjugates exhibited an emission peak at 518 nm, which was stable over time. A fluorescence sandwich-type immunoassay was used for the detection of PSA using FITC@SiO2-PBA-pAb. Alkaline hydrolysis of the sensing nanobioconjugates afforded enhanced sensitivity by releasing FITC molecules. In buffer samples, the fluorescent immunosensor exhibited a linear correlation range from 2.0 pg mL−1 to 50 ng mL−1. The linear range was from 2.0 pg mL−1 to 100 ng mL−1 in newborn calf serum (representing real samples). The limit of detection (LOD) was 8.25 fg mL−1 with a limit of quantification (LOQ) of 27.2 fg mL−1 in PBS (pH 7.4) after NaOH dissolution. A fluorescence immunosensor was used to detect PSA in spiked newborn calf serum with NaOH dissolution. It exhibited an LOD value of 33.0 fg mL−1 and LOQ value of 0.109 pg mL−1. The developed fluorescence immunosensor showed high selectivity and specificity for PSA. The detection of prostate-specific antigen in newborn calf serum samples exhibited no matrix interferences.

Abstract Image

用于超灵敏检测前列腺特异性抗原的抗体荧光素掺杂二氧化硅纳米生物共轭物
在此,我们报告了将抗前列腺特异性抗原多克隆抗体(pAb)生物共轭到掺杂荧光素的二氧化硅纳米颗粒上以检测前列腺特异性抗原(PSA)的方法。使用的荧光染料是荧光素-异硫氰酸酯(FITC),它与(3-氨基丙基)三乙氧基硅烷(APTES)反应生成 FITC-APTES 有机硅烷前体。将 FITC-APTES 与四乙氧基硅烷(TEOS)混合,形成含有 3% 和 6% 染料的荧光纳米二氧化硅颗粒(FITC@SiO2NPs)。二氧化硅纳米粒子可防止染料浸出,并促进荧光信号放大,用于检测 PSA。将苯硼酸(PBA)涂布在荧光纳米二氧化硅颗粒上,通过硼酸酯将抗体定向固定,形成 FITC@SiO2-PBA-pAb。这种荧光纳米二氧化硅生物共轭物在 518 纳米波长处显示出强烈的激发依赖性发射,并且长期稳定。利用 FITC@SiO2-PBA-pAb 的荧光夹心型免疫测定法检测 PSA。传感纳米生物共轭物经碱性水解后释放出 FITC 分子,从而提高了灵敏度。在缓冲液样品中,荧光免疫传感器的线性相关范围为 2.0 pg.mL-1 至 50 ng.mL-1。在新生小牛血清(代表真实样本)中,线性范围为 2.0 pg.mL-1 至 100 ng.mL-1。在 NaOH 溶解后的 PBS 缓冲液(pH 7.4)中,检测限(LOD)为 8.25 fg.mL-1,定量限(LOQ)为 27.2 fg.mL-1。研究了一种荧光免疫传感器,用于检测添加 NaOH 溶解物的新生小牛血清中的 PSA。LOD为33.0 fg.mL-1,LOQ为0.109 pg.mL-1。所开发的荧光免疫传感器对前列腺特异性抗原具有很高的选择性和特异性。在检测新生小牛血清样品中的前列腺特异性抗原时,未发现基质干扰。
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来源期刊
CiteScore
2.30
自引率
0.00%
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