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Biomimetic synthesis of all-inclusive organic-inorganic nanospheres for enhanced electrochemical immunoassay
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Luyuan Tiana, Yuxiao Mab, Ming Lia, Qiaorong Tanga, Luyang Miaoa, Bing Geng**, a, He Li*, a, c
Chinese Chemical Letters | 2020, 31(5) : 1159 - 1161
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Chinese Chemical Letters | 2020, 31(5): 1159-1161
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Biomimetic synthesis of all-inclusive organic-inorganic nanospheres for enhanced electrochemical immunoassay
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Luyuan Tiana, Yuxiao Mab, Ming Lia, Qiaorong Tanga, Luyang Miaoa, Bing Geng**, a, He Li*, a, c
Affiliations
  • a School of Chemistry and Chemical Engineering and Institute of Surface Analysis and Chemical Biology, University of Jinan, Ji'nan 250022, China
  • b The Hospital of University of Jinan, Ji'nan 250022, China
  • c College of Optoelectronics Technology, Chengdu University of Information Technology, Chengdu 610225, China
Published: 2020-05-15 doi: 10.1016/j.cclet.2019.10.007
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A friendly biomimetic process was adopted for the mild preparation of "all-inclusive" organic-inorganic nanospheres, which effectively integrate biorecognition function and signal amplification function. The resulted Ca3(PO4)2-Ab2-BSA nanospheres were employed as signal labels for enhancing detection of nuclear matrix protein 22 (NMP 22). The fabricated electrochemical immunosensor exhibited a linear range (0.08-77.00 U/mL) and an ultralow limit of detection (0.01 U/mL) towards NMP 22, which can be taken as a promising tool for clinical diagnosis of bladder cancer.

Nuclear matrix protein 22  /  Biomimetic process  /  Electrochemical immunosensor  /  Limit of detection
Luyuan Tian, Yuxiao Ma, Ming Li, Qiaorong Tang, Luyang Miao, Bing Geng, He Li. Biomimetic synthesis of all-inclusive organic-inorganic nanospheres for enhanced electrochemical immunoassay[J]. Chinese Chemical Letters, 2020 , 31 (5) : 1159 -1161 . DOI: 10.1016/j.cclet.2019.10.007
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Bladder cancer (BC) is the second most common genitourinary malignancy [1]. Clinical diagnosis of BC has great meaning for the effective treatment of BC, cystoscopy has been the clinical standard for the identification of bladder cancer, while the relative high cost and invasive detection manner will bring heavy burden for BC patients [2, 3]. Therefore, it is a high desire to develop sensitive, cost-effective and noninvasive methods for the detection of BC. Nuclear matrix protein 22 (NMP 22) as a tumor marker has been approved by US Food and Drug Administration for clinical diagnosis of BC and the clinical judgment value in urine was 10 U/mL, and if the value was higher than 10 U/mL, the detection results were positive [4].
Accordingly, some methods have been developed for sensitive detection of NMP 22, such as fluorescent, colorimetric [5], electrochemiluminescence [6], electrochemical [7-12]. Benefit from the advantages of low cost, friendly operation, rapid readout and satisfied sensitivity, electrochemical methodology has become a welcome method for tumor diagnosis beyond BC detection [13]. It is well worth to mention that some excited advancements have been made using nanomaterials to enhance the sensitivity of electrochemical immunoassay for protein biomarkers, which offer more accurate cancer diagnosis [14].
Recently, extensive efforts have been made to prepare "all-inclusive" or "all-in-one" nanomaterials for biomedical applications [15-19]. "All-in-one" nanomaterials can integrate diagnostic com-ponent and therapeutic component into one single unit, which will be an ideal platform for healthcare. Moreover, "all-in-one" nano-materials combining recognition function and signal amplification function do favor for sensing and biosensing applications.
Inspired by the mentioned work, we proposed a facile biomineralization method to prepare a novel kind of "all-inclusive" organic-inorganic nanospheres, Ca3(PO4)2-Ab2-BSA, as signal labels (Fig. 1A). Ca3(PO4)2-Ab2-BSA nanospheres possess the unique features: Ab2 will play biorecognition function for target analytes, BSA will diminish the non-specific adsorption and the inorganic component of Ca3(PO4)2 can supply plenty of phosphate ions to react with molybdate ions in acid medium to produce a large amount of redox-active molybdophosphates, which is the origin of the detection electrochemical signal. Much more produced molybdo-phosphates can significantly amplify the detection signal of the electrochemical immunoassay. Herein, we fabricated a "sandwich" type electrochemical immunosensor for bladder cancer using Ca3(PO4)2-Ab2-BSA as signal labels. As demonstrated in Fig. 1B, carboxylated single-wall carbon nanotubes (CNT) were used as a substrate material to increase conductivity. After the sandwich type immunoreactions were finished, the following reaction of "all-inclusive" Ca3(PO4)2-Ab2-BSA nanospheres with molybdate will generate electrochemical current which is relation to the concen-trations of NMP 22.
To our best knowledge, Ab2, as protein, contain some acidic amino acid residues which can chelate calcium ions, and then providing nucleation sites [20]. Nanoscale calcium phosphate crystals were constantly generated around the periphery of the peptide through spontaneous nucleation, and according to the bricks-and-mortar self-assembly mechanism, organized spherical structures was formed [21]. The morphology of the as-synthesized Ca3(PO4)2-Ab2-BSA were recorded by transmission electron microscopy (TEM). As illustrated by the SEM image in Fig. 2a, the Ca3(PO4)2 nanoparticles showed spherical structure with a diameter around 200 nm. The TEM image (Fig. 2b) of the Ca3(PO4)2 also exhibited typical spherical structure. The XRD pattern (Fig. 2c) of the calcium phosphate nanospheres was well-indexed to Ca3(PO4)2 xH2O (JCPDS card No. 18-0303). The first decline before 200 C was contributed to the decreased of H2O and the content of H2O was about 7.2 wt%. And the organic components including BSA and Ab2 decomposed from 200 C to 800 C, the content was around 22.3 wt% and the residual substance was Ca3(PO4)2 (Fig. 2d).
A series of electrochemical immunosensors were developed for detecting different concentrations of NMP 22. Under the optimized conditions (Fig. S4 in Supporting information), the concentrations of NMP 22 were correlation with the SWV peak currents at 0.16 Ⅴ (Fig. 3a). Importantly, the electrochemical immunosensor presents a sensitive detection performance toward NMP 22 which has a relationship with the logarithm of the NMP 22 concentration, ranging from 0.08 U/mL to 77.00 U/mL. Moreover, the calibration curve of the developed immunosensor was I (μA) = 17.95 5.40logCNMP 22 (U/mL) (R2 = 0.994), as shown in Fig. 3b and the detect limit was 0.01 U/mL (S/N = 3).
Meanwhile, the established electrochemical immunosensor possessed excellent reproducibility and good stability for the detection of NMP 22 (Fig. S5 in Supporting information). In order to assess the application potential and reliability of this immunosensor for real sample analysis, five urine samples (Numbering them as 1, 2, 3, 4, 5. Among of them, samples 1, 4 and 5 are negative, the other two are positive) diluted 100 times were conducted for further evaluation. As shown in Table S1 (Supporting information), the results matched well with the actual states of samples. Thus, the immunosensor is promising for clinical determination of NMP 22.
In summary, a satisfied sandwich electrochemical immunosen-sor was developed for NMP 22 detection. The good performance of the immunosensor was attributed to the promoted electrons transfer owing to the good conductivity of CNT and the amplified detection current signal by the transformation from Ca3(PO4)2 to redox-active molybdophosphates.
Declaration of competing interest
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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgments
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This work was supported by the Natural Science Foundation of Shandong Province (No. ZR2017MB017). We also gratefully acknowledge the support from the One-Thousand-Talents Scheme in Sichuan Province.
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Appendix
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Year 2020 volume 31 Issue 5
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Article Info
doi: 10.1016/j.cclet.2019.10.007
  • Receive Date：2019-07-25
  • Online Date：2026-01-31
  • Published：2020-05-15
Article Data
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History
  • Received：2019-07-25
  • Revised：2019-10-07
  • Accepted：2019-10-09
Affiliations
    a School of Chemistry and Chemical Engineering and Institute of Surface Analysis and Chemical Biology, University of Jinan, Ji'nan 250022, China
    b The Hospital of University of Jinan, Ji'nan 250022, China
    c College of Optoelectronics Technology, Chengdu University of Information Technology, Chengdu 610225, China
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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