Mouse protein arrays from a TH1 cell cDNA library for antibody screening and serum profiling

Gutjahr, Claudia; Murphy, Derek; Lueking, Angelika; Koenig, Andrea; Janitz, Michael; O’Brien, John; Korn, Bernhard; Horn, Sabine; Lehrach, Hans; Cahill, Dolores J.

doi:10.1016/j.ygeno.2004.11.005

Cited by 23 publications

(10 citation statements)

References 45 publications

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“…In this method, antibody-antigen interactions have been studied to identify autoantibodies from patients with autoimmune diseases and cancers such as colorectal, breast, ovarian, stomach, lung, and prostate cancer, and HCC [56,60,62,93,[121][122][123][124][125]. Because the microarray technology provides multiplexed analyses of thousands of proteins, this method permits highthroughput identification of TAA signatures for the development of cancer diagnostics and vaccines [126,127].…”

Section: Protein Microarraymentioning

confidence: 99%

Serum autoantibodies as biomarkers for early cancer detection

et al. 2009

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Autoantibodies against autologus tumor‐associated antigens have been detected in the asymptomatic stage of cancer and can thus serve as biomarkers for early cancer diagnosis. Moreover, because autoantibodies are found in sera, they can be screened easily using a noninvasive approach. Consequently, many studies have been initiated to identify novel autoantibodies relevant to various cancer types. To facilitate autoantibody discovery, approaches that allow the simultaneous identification of multiple autoantibodies are preferred. Five such techniques – SEREX, phage display, protein microarray, SERPA and MAPPing – are discussed here. In the second part of this review, we discussed autoantibodies found in the five most common cancers (lung, breast, colorectal, stomach and liver). The discovery of panels of tumor‐associated antigens and autoantibody signatures with high sensitivity and specificity would aid in the development of diagnostics, prognostics and therapeutics for cancer patients.

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Section: Protein Microarraymentioning

confidence: 99%

Serum autoantibodies as biomarkers for early cancer detection

et al. 2009

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“…With the development of etiological detection technology, microarrays have rapidly expanded into the area of biological research, including gene expression, signal transection, genome mismatch scanning, and protein trafficking. Among the many types of microarrays, the protein microarray offers an opportunity to study the full spectrum of protein attributes in a parallel, miniaturized, and automated fashion representing a significant shift from the traditional “one protein at a time” methods [58], [59]. Furthermore, the method not only aids in improved diagnosis, but also identifies autoantibody signatures that may represent disease subgroups, early diagnostics [60], [61] and facilitates the analysis of the outcome of vaccine trials [62].…”

Section: Discussionmentioning

confidence: 99%

A Protein Microarray for the Rapid Screening of Patients Suspected of Infection with Various Food-Borne Helminthiases

Chen

et al. 2012

PLoS Negl Trop Dis

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BackgroundFood-borne helminthiases (FBHs) have become increasingly important due to frequent occurrence and worldwide distribution. There is increasing demand for developing more sensitive, high-throughput techniques for the simultaneous detection of multiple parasitic diseases due to limitations in differential clinical diagnosis of FBHs with similar symptoms. These infections are difficult to diagnose correctly by conventional diagnostic approaches including serological approaches.Methodology/Principal FindingsIn this study, antigens obtained from 5 parasite species, namely Cysticercus cellulosae, Angiostrongylus cantonensis, Paragonimus westermani, Trichinella spiralis and Spirometra sp., were semi-purified after immunoblotting. Sera from 365 human cases of helminthiasis and 80 healthy individuals were assayed with semi-purified antigens by both a protein microarray and the enzyme-linked immunosorbent assay (ELISA). The sensitivity, specificity and simplicity of each test for the end-user were evaluated. The specificity of the tests ranged from 97.0% (95% confidence interval (CI): 95.3–98.7%) to 100.0% (95% CI: 100.0%) in the protein microarray and from 97.7% (95% CI: 96.2–99.2%) to 100.0% (95% CI: 100.0%) in ELISA. The sensitivity varied from 85.7% (95% CI: 75.1–96.3%) to 92.1% (95% CI: 83.5–100.0%) in the protein microarray, while the corresponding values for ELISA were 82.0% (95% CI: 71.4–92.6%) to 92.1% (95% CI: 83.5–100.0%). Furthermore, the Youden index spanned from 0.83 to 0.92 in the protein microarray and from 0.80 to 0.92 in ELISA. For each parasite, the Youden index from the protein microarray was often slightly higher than the one from ELISA even though the same antigen was used.Conclusions/SignificanceThe protein microarray platform is a convenient, versatile, high-throughput method that can easily be adapted to massive FBH screening.

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“…The amount of serum required to develop an array containing over 10,000 features can be the same as that required to develop a single feature in a well of a 96-well plate. Genetix microarrayers have been used for a range of such applications [4][5][6][7][8] . There are some cases, however, in which microwells have an advantage in that they can be easier to handle with existing instrumentation.…”

Section: Application Notesmentioning

confidence: 99%

The QArray series: new answers for protein arrays

Burke¹,

Munro²,

Mann³

2006

Nat Methods

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Screening tens to hundreds of proteins for enzymatic activity or protein interactions can now be performed in a quantitative and economical manner using protein microarrays. Recent validated examples include assays for human protein kinases and protein interactions with p53. Here we describe the QArray series of instruments for the printing and assaying of such arrays. Of particular note is printing of arrays into microwell plates, a methodology that combines the benefits of arrays and the convenience of a conventional assay format.Genetix markets a number of products for protein arraying. All three arrayers, QArray Mini , QArray2 or QArray Max use pins for spotting proteins onto surfaces (Fig. 1). These give less than 3% variation under standard conditions, and work to date indicates that the spotted proteins maintain their activity and are not denatured or inactivated by contact with the tungsten pins or various array surfaces.In the early development of protein arrays, much controversy surrounded whether the arrays should be made by contact printing with pins or by noncontact methods. Pins are now accepted as the industry standard. The difficulty with noncontact systems is that accumulation of static charge can cause inaccurate deposition of the spots (Fig. 2), and the expensive and delicate arraying pipettes have to be cleaned between the application of each protein to an array. In the case when many different proteins are arrayed on a relatively small number of slides (<100), this adds substantially to the time required to make the arrays.Arraying DNA onto surfaces usually requires either modification of the surface to create a three-dimensional surface or chemical modification (amide, aldehyde or epoxy groups) to couple the oligonucleotides or cDNAs to the array substrate.Protein arrays are created by printing spots of protein (for example, antibodies, recombinant proteins, peptides) on a solid surface such as a glass slide coated with a protein-binding chemistry. For such arrays, it is typical to modify the surface with molecules such as nickel or streptavidin to bind His-tagged or biotin-labeled proteins, respectively. This, of course, limits the range of proteins to be arrayed and requires specific surfaces for specific protein constructs. The production of proteins in bacterial or human cell culture systems are well-established procedures, and such proteins can be expressed with specific fusion tags for immobilization to appropriately coated solid surfaces. For nontagged proteins, the same surfaces used for DNA arrays such as amine, aldehyde or epoxy chemistries can be used for the capture of proteins onto the array.Two factors that are important for arraying proteins are humidity and temperature. These can be regulated in the Genetix QArray series of arrayers.Examples of proteins that have been successfully arrayed on chips are kinases and p53 (ref. 1). These are clearly important in cell signaling, and validate the approach of using arrays for investigating both the substrate specificity of mutants (...

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Mouse protein arrays from a TH1 cell cDNA library for antibody screening and serum profiling

Cited by 23 publications

References 45 publications

Serum autoantibodies as biomarkers for early cancer detection

Serum autoantibodies as biomarkers for early cancer detection

A Protein Microarray for the Rapid Screening of Patients Suspected of Infection with Various Food-Borne Helminthiases

The QArray series: new answers for protein arrays

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