Analytical Chemistry Chemical Cytometry Quantitates Superoxide

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Figure 3. Performance of the developed ELISA on different solid support matrixes. Three categories of the solid support were chosen on the basis of their chemical properties, namely polystyrene (PS) which is hydrophobic (white circles); polymethyl methacrylate (PMMA), a hydrophilic polymer (black circles); and polycyclo-olefin polymer, trade name Zeonex, which is a neutral matrix (inverted black triangles). The error bars represent standard deviations. may be a factor in their reduced sensitivities. 14 The sensitivity and specificity of any immunoassay is dependent on the antigen capture efficiency of an antibody which is subsequently governed by the nature of antibody 15,16 and accounts for the different assay sensitivities of the commercially available kits employing different type of antibodies that are monoclonal or polyclonal at the capture stage in ELISA. Three different types of commercially relevant substrates, polystyrene (PS) (hydrophobic), polymethyl methacrylate (PMMA) (hydrophilic), and cyclo-olefin polymer (Zeonex) (inert), were used in the modified microtiter plate format. The HFA assay was performed on these matrixes in the modified microtiter plate format. The HFA assays performed on different solid substrates could not be directly compared because of their different thick- (14) Wild, D. The Immunoassay Handbook, 3rd ed.; Elsevier: Oxford, 2005. (15) Lacy, A.; Dunne, L.; Fitzpatrick, B.; Daly, S.; Keating, G.; Baxter, A.; Hearty, S.; O’Kennedy, R. J. AOAC Int. 2006, 89 (3), 884–892. (16) Byrne, B.; Stack, E.; Gilmartin, N.; O’Kennedy, R. Sensors 2009, 9, 4407– 4445. 7052 <strong>Analytical</strong> <strong>Chemistry</strong>, Vol. 82, No. 16, August 15, 2010 nesses. Theoretically, the optical path length contributes toward the absorbance (A). Therefore, a thick solid support increases the path length and, hence, significantly changes the absorbance. However, the generic nature of the developed ELISA procedure was successfully demonstrated (Figure 3). The HFA assay performed on the Zeonex support, which is an inert cyclic poly olefin derivative, had an analytical sensitivity of 19.5 pg/mL, while for the assay performed on PS and PMMA, it was 78 pg/mL. The analytical sensitivities for the HFA assay and the dose-response, which determines the slope-dependent linearity range of the assay, of HFA-anti-HFA for all the three solid supports used in this study suggests that this modified protocol increases the overall assay sensitivity and is not confined to the nature of the solid support used for capturing antibody. CONCLUSIONS A rapid sandwich ELISA procedure was developed for the highly sensitive detection of HFA/AHSG. It was based on the covalent immobilization of anti-HFA capture antibody on a 3-APTES-functionalized microtiter plate. The developed ELISA has comparatively better analytical sensitivity (39 pg/mL) and less variability in interday and intraday assay repeats than the conventional format, which has an analytical sensitivity of 624 pg/ mL. The surface chemistry developed for this study is generic and can be employed to activate polymer matrixes irrespective of their chemical nature as demonstrated on PS (hydrophobic), PMMA (hydrophilic), and polycyclo-olefin (inert). Therefore, this strategy can also be used for rapid assay development on various commercially relevant substrates and the screening of substrates for particular biosensor/diagnostic applications. ACKNOWLEDGMENT C.K.D. and S.K.V. contributed equally to this work. We acknowledge Bristol Myers Squibb (BMS), Syracuse, USA, and Industrial Development Agency, Ireland, for the financial support under the CBAS Project Code 116294. SUPPORTING INFORMATION AVAILABLE Additional information as noted in text. This material is available free of charge via the Internet at http://pubs.acs.org. Received for review May 21, 2010. Accepted July 7, 2010. AC101339Q
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Anal. Chem. 2010, 82, 6745-6750 Let
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purchased from Invitrogen-Molecular
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Figure 3. Electropherograms of TPP-
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Anal. Chem. 2010, 82, 6751-6755 Res
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(pH 8.0) with cysteine and cystamin
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Figure 4. Ion mobility mass spectra
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GOx glucose + O298 gluconic acid +
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coater at 2000 rpm for 20 s, and th
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Figure 5. Comparison of cyclic volt
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in channels with either no grooves
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indicators of atmospheric processin
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Figure 1. GC/MS total ion chromatog
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Table 2. Concentrations and Stable
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on a substrate are preferred. 20-24
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Figure 4. SERS analysis of NAADP co
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Anal. Chem. 2010, 82, 6775-6781 Hig
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tion, 2 µL of proprionaldehyde wer
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Figure 4. Analysis of 2a by HPLC-MS
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eaction of 1a with PBH can be condu
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Numerous references had demonstrate
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Figure 1. TEM images of the prepare
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Figure 4. Schematic representation
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esult in a big SPR signal change wi
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also reduces chemical noise, which
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Table 1. Extraction Yields, Liquid
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scanning of AMPP amides of the anal
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Anal. Chem. 2010, 82, 6797-6806 δ
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Figure 1. Schematic view of the pre
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from the specimen and enclosed in a
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Figure 4. (A) IRMS mass-44 chromato
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Table 3. Ambient Measurement Result
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Anal. Chem. 2010, 82, 6807-6813 Dir
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Polymerase (1 U per sample). Reacti
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of which was constant for all ampli
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analytically useful signals at less
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carbon black and RP-C18 for the ext
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solution in an equal volume, and 1
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Table 2. Concentrations and Ratios
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Anal. Chem. 2010, 82, 6821-6829 Mac
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mg/mL protein, followed by separati
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Figure 2. Productivity of SEQUST an
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Figure 4. High-resolution MS/MS spe
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Figure 6. Characterization of PSMs
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containing T-T mismatches. 23 Based
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Figure 1. Extinction spectra of sol
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Figure 3. (A) The value of Ex650 nm
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Figure 5. Extinction spectra and co
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wished to explore the dehydration o
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constant medium for separation; we
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Figure 2. Standards of (Pi)n, n ) 1
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Figure 5. Quantitative calibration
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Anal. Chem. 2010, 82, 6847-6853 Met
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Figure 1. 226 Ra spectrum by liquid
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Table 1. Counting Properties and De
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Table 3. Analysis of 226 Ra in Sedi
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mercial microarray scanner and fabr
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Figure 2. Optical transmission meas
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Figure 5. Volcano plots detailing t
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data as well. The 41 genes in Table
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corresponding compound if its chemi
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Figure 1. Schematic illustration of
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Table 1. Absolute Quantification Re
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ment. Therefore, the long-time drea
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Figure 1. Chemically actuated micro
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Figure 2. Influence of a surfactant
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Figure 5. Device to eject and mix s
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Anal. Chem. 2010, 82, 6877-6886 Imm
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NaCl, phosphate buffer saline (PBS)
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Figure 2. Product ion mass spectra
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-70 °C resolved the problem, givin
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Table 1. Intraday Precision and Acc
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Anal. Chem. 2010, 82, 6887-6894 Fer
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Figure 1. Infrared spectra of (A) u
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Figure 3. Cyclic voltammograms obta
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Figure 6. Calculated charge from ch
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Anal. Chem. 2010, 82, 6895-6903 Ele
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Table 1. Chemical Structure, pKa Va
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pH with a tilted baseline (Figure 1
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Table 2. Linearity and Detection Li
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ascorbic acid (AA), uric acid (UA),
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the multielement capabilities, the
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RESULTS AND DISCUSSION Sulfur Detec
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Table 2. Molecular Properties and C
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Anal. Chem. 2010, 82, 6911-6918 Dir
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dilution and hybridization buffer.
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solution under appropriate incubati
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Figure 4. Standardization curve for
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Anal. Chem. 2010, 82, 6919-6925 Ele
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the ×10 objective, to have a large
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Figure 2. With a suitable removal o
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the NB signal in a much better foot
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Scheme 1. Reactions of Selenium Rea
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Figure 2. (a) ESI-MS spectrum showi
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Figure 4. (a) ESI-MS spectrum showi
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Anal. Chem. 2010, 82, 6933-6939 Dif
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trode 28 by a finite element using
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Figure 4. Comparison between simula
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Figure 6. Comparison between simula
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educed in the vicinity of double bo
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Figure 2. Normalized product ion ab
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Figure 4. EID (a) and IRMPD (b) of
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Anal. Chem. 2010, 82, 6947-6957 Ide
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Figure 1. Schematic flowchart showi
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difference, ppm compound Table 1. I
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isoforms, its successful use, in th
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Figure 5. Extracted ion current ESI
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m/z 1172.935 was observed for Ser14
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linked products via affinity tags.
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Scheme 2. Fragmentation Mechanism o
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Figure 1. (A) ESI-LTQ-CID-MS 2 prod
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Figure 3. (A) ESI-LTQ-CID-MS 2 prod
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Figure 5. (A) MALDI-TOF/TOF product
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Anal. Chem. 2010, 82, 6969-6975 Ana
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Figure 3. Equilibrium response as a
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Figure 6. The average measured resp
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for the fill time, and we find that
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nucleotide tails. 3-5 Thus, the amo
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allow the use of higher aptamer con
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quent ligation of the aptamers afte
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Anal. Chem. 2010, 82, 6983-6990 Imp
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Figure 2. Configuration editor wind
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Figure 4. Dependencies between volu
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Since the time for liquid expulsion
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Anal. Chem. 2010, 82, 6991-6999 Int
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Figure 1. Representation of the Car
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Table 2. Capillary Electrophoresis
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Figure 4. (A) Typical raw electroph
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field. DNA profiles were delivered
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Page 261 and 262: Figure 5. Relevant score plots and
Page 263 and 264: CONCLUSIONS In this paper we have i
Page 265 and 266: electroactive-species loaded liposo
Page 267 and 268: Figure 2. Qdot-based FLFTS response
Page 269 and 270: Figure 6. Fluorescence imaging of Q
Page 271 and 272: Anal. Chem. 2010, 82, 7015-7020 Sel
Page 273 and 274: Table 1. Effect of 1 D-LC Condition
Page 275 and 276: Figure 3. Peak-production rate vers
Page 277 and 278: Anal. Chem. 2010, 82, 7021-7026 Cel
Page 279 and 280: luciferase (T7 control vector) was
Page 281 and 282: Figure 3. Inhibitory effects of luc
Page 283 and 284: Anal. Chem. 2010, 82, 7027-7034 Pat
Page 285 and 286: Electrochemistry. Electrochemical m
Page 287 and 288: Figure 2. SEM images of Au substrat
Page 289 and 290: Table 3. Film Thickness Measurement
Page 291 and 292: Anal. Chem. 2010, 82, 7035-7043 Qua
Page 293 and 294: Figure 1. (A) FL spectra of BSPOTPE
Page 295 and 296: Figure 4. (A) Variation in the FL i
Page 297 and 298: Figure 6. (A) FL spectra of BSPOTPE
Page 299 and 300: Scheme 1. Proposed Mechanism for Fl
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Page 303 and 304: Figure 2. Free fluorophores do not
Page 305 and 306: Anal. Chem. 2010, 82, 7049-7052 Dev
Page 307: Figure 2. Comparative analysis of d
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