Detection of Multiple Proteins on One Spot by Laser Ablation Inductively Coupled Plasma Mass Spectrometry and Application to Immuno- Microarray with Element-Tagged Antibodies

Hu, Shenghong; Zhang, Sichun; Hu, Zhaochu; Xing, Zhi; Zhang, Xinrong

doi:10.1021/ac061269p

Cited by 186 publications

(114 citation statements)

References 30 publications

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“…It provides an alternative strategy to count the number of OSHs and OSOSO involved, offering valuable information for proteomic studies. Such information is not only important to protein chemistry, but also crucial to the quantitative analysis of peptides and/or proteins when a hard ionization source mass spectrometry, such as inductively coupled plasma mass spectrometry (ICP-MS) [18,19], is applied to overcome the relatively low ionization ability of ESI-MS for labeled peptides and/or proteins. Both quantification of a protein and locating OSHs and OSOSO in a protein with ESI-MS together with ICP-MS using RHg ϩ as a tag will be the subjects of future studies.…”

Section: Resultsmentioning

confidence: 99%

Counting sulfhydryls and disulfide bonds in peptides and proteins using mercurial ions as an MS-tag

Guo

Chen

Yang

et al. 2008

J. Am. Soc. Mass Spectrom.

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Organic mercurial compounds are the most specific and sensitive reagents for reaction with the sulfhydryl groups (OSHs) in peptides and proteins because of the strong mercury-sulfur affinity. Using the monofunctional organic mercury ion RHg ϩ as a mass spectrometry (MS)-tag has the advantages of reacting with one sulfhydryl group, offering definite mass shift, and especially stable and characteristic nonradioactive isotopic distribution. Mass spectrometric analysis of derivatized sulfhydryls in peptides/proteins is thus an alternative for precisely counting the number of sulfhydryl groups and disulfide bonds (OSOSO). Here the tags used include monomethylmercury chloride, monoethylmercury chloride, and 4-(hydroxymercuri) benzoic acid. The feasibility of this strategy is demonstrated using HPLC/ESI-MS to count OSHs and OSOSO in model peptides/proteins, i.e., glutathione, phytochelatins, lysozyme and ␤-lactoglobulin, which contain increasing OSHs and various OSOSO linkages. (J Am

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Section: Resultsmentioning

confidence: 99%

Counting sulfhydryls and disulfide bonds in peptides and proteins using mercurial ions as an MS-tag

Guo

Chen

Yang

et al. 2008

J. Am. Soc. Mass Spectrom.

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“…The nanoparticles tagged on biological molecules could be detected by ICP-MS, which has been demonstrated by Baranov et al [20], Merkoci et al [21], and our group [22,23] in recent years. An important advantage of ICP-MS for nanoparticle detection is that high sensitivity could be obtained because of large quantities of detectable atoms in each nanoparticle tag.…”

mentioning

confidence: 99%

A new strategy for highly sensitive immunoassay based on single-particle mode detection by inductively coupled plasma mass spectrometry

Liu

Zhang

et al. 2009

J. Am. Soc. Mass Spectrom.

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A highly sensitive immunoassay is proposed based on time-resolved inductively coupled plasma mass spectrometry with nanoparticles as tags to antibody. Instead of using traditional integral mode detection, the transient signals induced by the flash of ions in the plasma torch from the ionization of nanoparticles tagged on antibody were recorded in a time-resolved mode. Since, under certain conditions, the frequency of transient signals is directly correlated to the concentration of nanoparticle tags, the concentration of nanoparticle-tagged antibodies can be quantified by the frequency of transient signals. With the present instrument setup, gold nanoparticle (Au-NP) tags, as small as about 15 nm in diameter, can be detected. This protocol is evaluated for a competitive immunoassay and the linear range for ␣-fetoprotein is 0.016 -6.8 g/L (between 20 and 80% inhibition). The limit of quantification is 0.016 g/L (20% inhibition, IC 20 ) with a relative standard deviation of 4.2% (20% inhibition, 4 replicates) for ␣-fetoprotein. The present strategy provides a sensitive readout method for nanoparticle tags, which is quite promising for numerous applications in immunoassay, DNA hybridization, and other biological analyses. (J Am Soc Mass

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“…Label-free immunoassays are difficult to develop because of the lack of a detectable protein signal or a signal too weak to quantify the trace amount of analytes (14 ). Labeled probe methods such as multilabel and spatially resolved assays can offer amplified detection signals for multianalyte immunoassays.…”

mentioning

confidence: 99%

A Disposable Multianalyte Electrochemical Immunosensor Array for Automated Simultaneous Determination of Tumor Markers

et al. 2007

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Background: Automated and convenient multianalyte detection with high throughput is increasingly needed in clinical diagnosis. We developed a disposable 4-by-2 array for programmed simultaneous amperometric immunoassay of 4 tumor markers. Methods: We used a screen-printed technique, 1-step immobilization method, and flow injection technique. We immobilized carcinoembryonic antigen, ␣-fetoprotein, ␤-human choriogonadotropin, and carcinoma antigen 125 as model analytes in a redox mediator-grafted, biopolymer-modified, screen-printed carbon electrode array to capture corresponding horseradish peroxidaselabeled antibodies in competitive immunoreactions. The simultaneous multianalyte immunoassay was automatically carried out to amperometrically monitor the mediator-catalyzed enzymatic response to hydrogen peroxide, which decreased in proportion to the concentrations of analytes in samples. Results: The multianalyte immunosensor array had a throughput of 60 samples/h and allowed simultaneous detection of carcinoembryonic antigen, ␣-fetoprotein, ␤-human choriogonadotropin, and carcinoma antigen 125 in clinical serum samples with concentrations up to 188 g/L, 250 g/L, 266 IU/L, and 334 kIU/L, respectively. The detection limits (limits of the blank, mean of blank plus 3 SD) were 1.1 g/L, 1.7 g/L, 1.2 IU/L, and 1.7 kIU/L. The inter-and intraassay imprecision (CVs) of the immunosensor arrays were <7.8% and <9.0%, respectively. The immunosensor arrays were stable for 28 days.

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Detection of Multiple Proteins on One Spot by Laser Ablation Inductively Coupled Plasma Mass Spectrometry and Application to Immuno- Microarray with Element-Tagged Antibodies

Cited by 186 publications

References 30 publications

Counting sulfhydryls and disulfide bonds in peptides and proteins using mercurial ions as an MS-tag

Counting sulfhydryls and disulfide bonds in peptides and proteins using mercurial ions as an MS-tag

A new strategy for highly sensitive immunoassay based on single-particle mode detection by inductively coupled plasma mass spectrometry

A Disposable Multianalyte Electrochemical Immunosensor Array for Automated Simultaneous Determination of Tumor Markers

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