SERS quantitative detection of trace human chorionic gonadotropin using a label‐free Victoria blue B as probe in the aggregated immunonanogold sol substrate

Nanogold particles (NG) were modified by anti‐rabbit antibody (RAb) against human chorionic gonadotropin to obtain an immunonanogold probe (ING). In pH 7.0 Na₂HPO₄‐citrate buffer solution containing KCl, ING probes formed large aggregates in which Victoria blue B (VBB) molecules were adsorbed on the surface and which exhibited strong surface‐enhanced Raman scattering (SERS) at a peak of 1612 cm^–1. After addition of human chorionic gonadotropin (hCG) an immune reaction with the ING probe occurred to form dispersive ING–hCG complexes with non‐SERS activity that led to a decreased SERS peak at 1612 cm^–1. The decreased SERS intensity was linear to the concentration of hCG over 2.4–73.2 ng/mL. The ING reaction was studied in detail by SERS, scanning electron microscope (SEM), resonance Rayleigh scattering (RRS), surface plasmon resonance (SPR) absorption and laser scattering techniques. SERS quenching was observed and discussed. Copyright © 2014 John Wiley & Sons, Ltd.     

Resonance scattering spectral detection of ultratrace Hg(II) using herring sperm DNA modified nanogold probe as catalyst

Herring sperm DNA (hsDNA) was used to modify 10 nm nanogold to obtain a resonance scattering (RS) probe (AuhsDNA) for detection of Hg²⁺. In the presence of salt, Hg²⁺ interacts with AuhsDNA to form stable Hg²⁺–hsDNA complexes, and releases nanogold particles to form larger nanogold clusters that can be removed by membrane filtration. The excess AuhsDNA in the filtrate solution exhibits a catalytic effect on the reaction between hydroxylamine (NH₂OH) and Cu(II). The excess AuhsDNA decreased with the addition of Hg²⁺, which led the RS intensity at 602 nm to decrease. The decreased RS intensity (Δl_{602 nm}) had a linear response to Hg²⁺ concentration in the range of 0.4–400 nM, with a detection limit of 0.2 nM Hg²⁺. This RS method was applied for the detection of Hg²⁺ in water samples, with sensitivity, selectivity and simplicity. Copyright © 2009 John Wiley & Sons, Ltd.     

Highly Sensitive Resonance Scattering Detection of DNA Hybridization Using Aptamer-Modified Gold Nanopaticle as Catalyst

Gold nanoparticle particles in size of 10 nm were used to label the thiol-modified single-stranded DNA aptamer (SH-ssDNA) to obtain an aptamer-modified gold nanoparticle probe (AussDNA) for target DNA (tDNA). In pH 7.4 NaH₂PO₄–Na₂HPO₄ buffer solution, the hybridization reaction between AussDNA and tDNA took place to form larger aptamer-modified gold nanoparticle cluster complex. The excess aptamer-modified gold nanoparticle probe in the supernatant solutions was obtained by centrifuging and can be used as nanocatalyst for the 0.276 mmol/L CuSO₄-65.4 mmol/L potassium-sodium tartrate-0.37 mmol/L glucose system at 70 °C. The cubic Cu₂O particles generated by the nanocatalytic reducing exhibit a strong resonance scattering (RS) peak at 620 nm. In the selected conditions, the RS intensity at 620 nm decreased with addition of tDNA, and the decreased intensity ΔI _{620 nm} is proportional to tDNA concentration (C _tDNA) from 0.12 to 72 pM, with regress equation of ΔI _620 nm = 1.29C _tDNA + 4.05, correlation coefficient of 0.9917, and detection limit of 0.084 pM tDNA.     

Resonance Scattering Spectral Detection of Trace Pb<Superscript>2+</Superscript> Using Aptamer-Modified AuPd Nanoalloy as Probe

The resonance scattering spectral probe for Pb²⁺ was obtained using aptamer-modified AuPd Nanoalloy. In the pH 7.0 Na₂HPO₄–NaH₂PO₄ buffer solution, the aptamer interacted with AuPd nanoalloy particles to form stable aptamer-AuPd nanoalloy probe for Pb²⁺ that is stable in high concentration of salt. The probe combined with Pb²⁺ ions to form a G-quadruplex and to release AuPd nanoalloy particles that aggregate to form big particles which led the resonance scattering (RS) intensity enhancing. The reaction solution was filtered by 0.15 μm membrane to obtain the filtration containing aptamer-AuPd nanoalloy probe that has strong catalytic effect on the electrodeless nickel particle plating reaction between Ni(II) and PO₂³⁻ that exhibited a strong RS peak at 508 nm. The RS intensity at 508 nm decreased when the Pb²⁺ concentration increased. The decreased intensity (ΔI _508nm) is linear to the concentration of 0.08–42 nM Pb²⁺, with regress equation of DI_508nm = 16.3 c + 1.5 \Delta {I_{{5}0{\rm{8nm}}}} = {16}.{3}\,c + {1}.{5} , correlation coefficient of 0.9965, and detection limit of 0.04 nM Pb²⁺. The RS assay was applied to the analysis of Pb²⁺ in wastewater, with satisfactory results.     

Rapid assay of trace immunoglobulin M by a new immunonanogold resonance scattering spectral probe

Nanogold, 8 nm in size, was used to label goat antihuman immunoglobulin M (GIgM) to obtain a new immunonanogold resonance scattering (RS) probe (Au-GIgG) for quantitation of trace immunoglobulin M (IgM). The Au-GIgG combined with IgM to form nanogold-labeled immunocomplex causes the RS intensity at 580 nm to be enhanced, in pH 4.49 KH(2)PO( 4)-Na(2)HPO(4) buffer and in the presence of polyethylene glycol 6000. The enhanced RS intensity at 580 nm (DeltaI(580 nm)) is proportional to the IgM concentration in the range of 1.5 to 2000 ng/mL, with a lower detection limit of 0.98 ng/mL. The immunonanogold RS assay was used to assay IgM in serum samples, with sensitivity, selectivity, and simplicity.     

A new and sensitive catalytic resonance scattering spectral assay for the detection of laccase using guaiacol as substrate

In pH 4.0 succinic acid-sodium hydroxide buffer solution, laccase catalyzed the oxidization of guaiacol substrate to form red particles, which exhibited a strong resonance scattering (RS) peak at 590 nm. Under the chosen conditions, as the laccase increased, the RS intensity (ΔI) increased linearly. The ΔI was proportional to laccase activity in the range of 0.10-1.2 U/mL, with a regression equation of ΔI = 734.0 U(laccase) - 9.7, and a detection limit of 0.05 U/mL. This RS method was applied to the detection of laccase activity in real samples, and the results were agreement with those from spectrophotometry.     

A Simple and Rapid Resonance Scattering Spectral Method for Detection of Trace Hg<Superscript>2+</Superscript> Using Aptamer-Nanogold as Probe

Single-strand deoxyribonucleic acid (ssDNA) were used to modified nanogold particle to obtain a aptamer-nanogold probe (NGssDNA) for Hg(II). The probe is not aggregated in high concentration of NaCl. In the pH 7.0 Na₂HPO₄-NaH₂PO₄ buffer solution and in the presence of high concentration of NaCl, NGssDNA interact with Hg(II) to form stable double-strand T-Hg(II)-T mismatches and to release nanogold particles from the probe. The released nanogold particles aggregated to form bigger clusters which leaded the resonance scattering (RS) intensity at 540 nm enhanced linearly with the concentration of Hg²⁺ in the range of 0.39–1666.7 nM, with detection of 0.1 nM. This simple, rapid, and sensitive aptamer-nanogold RS assay was applied to determination of Hg²⁺ in wastewater, with satisfactory results.     

A Highly Sensitive Resonance Scattering Assay for Immunoglobulin M Using Ag(I)–Hydroquinone–Immunonanogold Catalytic Reaction

Ten-nanometer nanogold showed the strongest catalytic effect on the particle reaction between Ag(I) and hydroquinone to form nanosilver particles that exhibited the strongest resonance scattering (RS) peak at 350 nm. The enhanced RS intensity was linear to the nanogold concentration in the range of 30–5,700 nM Au. The nanogold was used to label goat antihuman immunoglobulin M (GIgM) to obtain an immunonanogold probe (AuGIgM) for immunoglobulin M (IgM). Based on the nanogold-labeled immunoreaction between IgM and AuGIgM, centrifugation, and AuGIgM–Ag(I)–hydroquinone nanocatalytic reaction, a highly sensitive and selective immunonanogold-catalytic Ag particle RS assay for 0.2–300 ng mL^?1 IgM was proposed, with a detection limit of 0.1 ng mL^?1. This assay was simple and sensitive and was applied to assay IgM in serum samples, with satisfactory results.     

A highly sensitive resonance scattering spectral assay for carcinoembryonic antigen using immunonanogold as probe and nanocatalyst of NH2OH�CCu(II) particle reaction

Nanogold of 10 nm was used to label carcinoembryonic antigen antibody (CEAAb) to prepare a probe (Au-CEAAb) for carcinoembryonic antigen (CEA). In a Na₂HPO₄–NaH₂PO₄ buffer solution of pH 6.8, CEA reacted with Au-CEAAb to form a big Au-CEAAb–CEA immunocomplex that can be removed by centrifugation. The unreacted Au-CEAAb in the centrifugal supernatant exhibited catalytic effect on the Cu₂O particle reaction, and the Cu₂O particles displayed a resonance scattering (RS) peak at 602 nm. When CEA increased, the RS intensity at 602 nm decreased, and the decreased RS intensity (ΔI _602 nm) was linear to CEA concentration (C _CEA) in the range of 0.02–12 ng mL⁻¹, with the regression equation of ΔI _602 nm = 27.1 C _CEA + 3.3, correlation coefficient of 0.9978 and detection limit of 3 pg mL⁻¹ CEA. The proposed method was applied to detect CEA in real samples, with satisfactory results.     

A selective resonance scattering assay for immunoglobulin G using Cu(II)-ascorbic acid-immunonanogold reaction

In sodium acetate–acetic acid buffer solution, Au, Ag, Pt, Pd, Fe₃O₄, and Cu₂O nanoparticles have catalytic enhancement effect on the reduction of Cu²⁺ by ascorbic acid to form large copper particles that exhibit a strong resonance scattering peak at 610 nm. Those nanocatalytic reactions were studied by the resonance scattering spectral technique, and smaller nanogold exhibited stronger catalytic enhancement effect in pH 4.2 sodium acetate–acetic acid buffer solution. The resonance scattering intensity at 610 nm increased linearly with the concentrations of 0.02 to 1.60, 0.040 to 1.20, and 0.12 to 4.70 nM nanogold in sizes of 5, 10, and 15 nm with detection limits of 0.010, 0.030, and 0.10 nM, respectively. An immunonanogold-catalytic resonance scattering bioassay was established, combining the immunonanogold-catalytic effect on CuSO₄–ascorbic acid reaction with the resonance scattering detection technique. As a model, 0.03 to 7.5 ng ml⁻¹ immunoglobulin G can be assayed by this immunonanogold-catalytic resonance scattering bioassay with a detection limit of 0.015 ng ml⁻¹.     

A sensitive and selective immuno‐nanogold resonance‐scattering spectral method for the determination of trace penicillin G

A sensitive and selective immuno‐nanogold resonance scattering spectral assay was developed for the determination of trace hapten penicillin G, based on the resonance scattering (RS) effect of the nanogold at 560 nm, and the nanogold‐labelled immunoreaction took place in pH 5.4 phosphate citric acid buffer solutions and in the presence of polythylene glycol (PEG). The nanogold‐labelled immunocomplex formed more and more with addition of penicillin G. The enhanced RS intensity at 560 nm ΔI_RS was linear to the penicillin G concentration in the range 7.5–1700 ng/mL, with a detection limit of 0.78 ng/mL. The results indicate that the immunonanogold‐labelled RS spectral assay has a high specificity and sensitivity for quantitative determination of penicillin G in raw milk samples. Copyright © 2008 John Wiley & Sons, Ltd.     

A sensitive enzyme-catalytic nanogold-resonance scattering spectral assay for alkaline phosphate

In pH 8.9 Tris-HCl buffer solutions, alkaline phosphatase (ALP) catalyzed the hydrolysis of ascorbic acid 2-phosphate (AAP) substrate to form ascorbic acid. Then H(3)PO(4) was added to stop the enzymatic reaction and HAuCl(4) was used to react with ascorbic acid to generate gold nanoparticles that exhibited a resonance scattering (RS) peak at 600 nm. Under the selected conditions, when the activity of ALP increased, the formed ascorbic acid and gold nanoparticles also increased. Thus, the RS intensity at 600 nm enhanced linearly. The linear range was 0.06-22 U/L, with a detection limit of 0.03 U/L. The ALP in serum was analyzed, and the results were in agreement with those of the fluorescence method.     

Catalytic effect of ReAu nanoalloy on the Te particle reaction and its application to resonance scattering spectral assay of CA125

ReAu nanoparticles with a molar ratio of 2:8 Re and Te nanoparticles were prepared by NaBH₄ reduction. In HCl medium at 65°C, ultratrace Re, Te and ReAu bimetallic nanoparticles strongly catalyzed the slow reaction between Sn(II) and Te(VI) to form Te particles, which exhibited the strongest resonance scattering (RS) peak at 782 nm. As the amount of nanocatalyst increased, the RS intensity at 782 nm (I_{782 nm}) increased linearly, and the increase in intensity ΔI_{782 nm} was linear to the ReAu, Re and Te concentrations in the ranges 0.07–9.0, 0.01–4.5 and 30–1200 nm , respectively. As a model, a ReAu immunonanoprobe catalytic Te–particle resonance scattering spectral (RSS) method was established for detection of CA125, using ReAu nanoparticle labeling CA125 antibody (CA125Ab) to obtain an immunonanoprobe (ReAuCA125Ab) for CA125. In pH 7.6 citric acid–Na₂HPO₄ buffer solution, ReAuCA125Ab aggregated nonspecifically. Upon addition of CA125, the immunonanoprobe reacted with it to form ReAuCA125Ab–CA125 dispersive immunocomplex in the solution. After the centrifugation, the supernatant containing the immunocomplex was used to catalyze the reaction of Te(VI)–Sn(II) to produce the Te particles that resulted in the I_{782 nm} increasing. The ΔI_{782 nm} was linear to CA125 concentration (C_CA125) in the range 0.1–240 mU/mL. The regression equation, correlation coefficient and detection limit were ΔI_{782 nm} = 1.61 C_CA125 + 1.5, 0.9978 and 0.02 mU/mL, respectively. The proposed method was applied to detect CA125 in serum samples, with satisfactory results. Copyright © 2010 John Wiley & Sons, Ltd.     

A Highly Sensitive Enzyme Catalytic Method for the Detection of Ethanol Based on Resonance Scattering Effect of Gold Particles

In pH 8.4 Tris–HCl buffer solutions, alcohol dehydrogenase catalyzed the reaction between ethanol and nicotinamide adenine dinucleotide to produce acetaldehyde. In the medium of HCl, acetaldehyde reduced HAuCl₄ to form gold particles that exhibited a strong resonance scattering (RS) peak at 600 nm. The RS peak increased with ethanol concentration. The increased RS intensity at 600 nm (ΔI _600 nm) was proportional to the ethanol concentration (C) from 0.068 to 10.2 mmol/L, with a regression equation of ΔI _600 nm?=?35.59?C?+?16.1, and a detection limit (3σ) of 3.2 μmol/L. This proposed method was applied to detect ethanol in saliva and plant cell culture medium samples, with satisfactory results.     

A Simple and Sensitive Resonance Scattering Spectral Assay for Detection of Melamine Using Aptamer-Modified Nanosilver Probe

Nanosilver of 10-nm size was prepared by the NaBH₄–sodium citrate procedure, and it was modified by a single-strand DNA (ssDNA) aptamer to fabricate an AgssDNA probe for melamine. The probe was stabile at pH 7.0 Na₂HPO₄–NaH₂PO₄ buffer solutions and in the presence of 25.0 mmol/L NaCl. Upon the addition of melamine, it interacted with the probe to aggregate big clusters, which led to the resonance scattering (RS) intensity at 470 nm increasing greatly. Under the selected conditions, the increased RS intensity (ΔI _470 nm) is linear to melamine concentration in the range of 6.31–378.4 μg/L, with a regression equation of DI_{470 nm} = 1.124c + 10.8 \Delta {I_{{47}0{\rm{ nm}}}} = {1}.{124}c + { 10}.{8} and a detection limit of 3.1 μg/L. The aptamer-modified nanosilver RS assay has been applied for the determination of melamine in milk, with satisfactory results.     

Resonance light scattering determination of trace bisphenol A with signal amplification by aptamer–nanogold catalysis

HAuCl₄ was reduced by sodium citrate to prepare 10 nm gold nanoparticles (AuNPs) that were modified by the bisphenol A aptamer (Apt) to obtain an aptamer–nanogold probe (Apt‐AuNP) for bisphenol A (BPA). The probes were aggregated nonspecifically to form large clusters, which showed a strong resonance light scattering (RLS) peak at 520 nm, under preparation conditions (pH 7.6 Na₂HPO₄‐NaH₂PO₄ buffer and ultrasonication). Upon addition of BPA, the probe reacted specifically to form dispersed BPA‐Apt‐AuNP conjugates that exhibited strong catalysis of the two particle reactions of glucose‐Cu(II) and hydrazine hydrochloride‐Cu(II) with a strong RLS peak at 360 nm and 510 nm respectively. When the BPA concentration increased, the RLS intensity at 360 nm and 510 nm increased respectively. Accordingly, two new and highly‐sensitive RLS methods were established for the detection of BPA, using the Apt‐AuNP catalytic amplification. Copyright © 2013 John Wiley & Sons, Ltd.     

A new and simple resonance Rayleigh scattering method for human serum albumin using graphite oxide as probe

Graphite oxide (GO) was prepared by the Hummer procedure, and can be dispersed to stable colloid solution by ultrasonic wave. The GO exhibited an absorption peak at 313 nm, and a resonance Rayleigh scattering (RRS) peak at 490 nm. In pH 4.6 HAc‐NaAc buffer solution, human serum albumin (HSA) combined with GO probe to form large HSA‐GO particles that caused the RRS peak increasing at 490 nm. The increased RRS intensity was linear to HSA concentration in the range 0.50–200 µg/mL. Thus, a new and simple RRS method was proposed for the determination of HSA in samples, with a recovery of 98.1–104%. Copyright © 2012 John Wiley & Sons, Ltd.     

Small-angle X-ray scattering study by synchrotron orbital radiation reveals that high molecular weight subunit of glutenin is a very anisotropic molecule.

Small-angle X-ray scattering of one high molecular weight (HMW) subunit of wheat glutenin was measured at protein concentration ranges from 1.0 to 10.0 mg/ml. The radius of gyration of whole particles, RO, in aq. 50% (v/v) 1-propanol and 0.1M acetic acid was 16.6 +/- 0.1nm and 22.8nm, respectively, and the corresponding radius of gyration of the cross-section, RC, was 2.82 +/- 0.02 nm and 2.23 +/- 0.01 nm, which indicate that the glutenin HMW subunit exists as very anisotropic particles in both solutions. The RO and RC values of the subunit, and the drastic decrease in scattered intensity at small angles that occurs in the acetic acid solution with relatively low protein concentration are completely explained in terms of rod-like molecules of the glutenin HMW subunit.     

A simple and sensitive resonance Rayleigh scattering method for determination of As(III) using aptamer‐modified nanogold as a probe

A simple and selective aptamer (ssDNA)‐modified nanogold probe (AussDNA) was prepared for the determination of trace As(III) in HEPES buffer solution (pH 8.2) containing 0.05 mol/L NaCl. The method coupled the aptamer reaction of AussDNA–As(III) and the resonance Rayleigh scattering (RRS) of nanogold aggregations at 278 nm. When the As(III) concentration increased, the RRS intensity at 278 nm increased to form more nanogold aggregation and a stable As(III)–ssDNA complex. Under selected conditions, the increased RRS intensity (ΔI) was linear to the concentration of As(III) in the range 3.8–230.4 ng/mL, with a detection limit of 1.9 ng/mL. This RRS method was applied to detect As(III) in water samples, with simplicity, sensitivity and selectivity. Copyright © 2013 John Wiley & Sons, Ltd.     

共振瑞利散射测试血清蛋白酸碱度影响的探讨

提出了一种基于共振瑞利散射（RRS）原理测量人体血清蛋白的新方法。在缓冲溶液的作用下,把配制好的人体血清蛋白稀释液按比例与四羧基酞菁锌混合,经过化学作用后在波长为400 nm左右蓝色波段强光照射下,散射出480 nm左右的共振瑞利散射光强信号。考察在不同pH对共振瑞利散射光强信号与混合物中的血清蛋白反应线性关系的影响。结果表明,pH在6.0～8.0范围内混合溶液共振瑞利散射光强信号与血清蛋白的线性关系良好。