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Proteome serological determination of tumor-associated antigens in melanoma.

Forgber M, Trefzer U, Sterry W, Walden P - PLoS ONE (2009)

Bottom Line: One of these antigens, galectin-3, has been related to various oncogenic processes including metastasis formation and invasiveness.Similarly, enolase has been found deregulated in different cancers.With at least 2 of 18 identified proteins implicated in oncogenic processes, the work confirms the potential of proteome-based antigen discovery to identify pathologically relevant proteins.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, Venerology and Allergy, Charité Universitätsmedizin Berlin, Humboldt University, Berlin, Germany.

ABSTRACT
Proteome serology may complement expression library-based approaches as strategy utilizing the patients' immune responses for the identification pathogenesis factors and potential targets for therapy and markers for diagnosis. Melanoma is a relatively immunogenic tumor and antigens recognized by melanoma-specific T cells have been extensively studied. The specificities of antibody responses to this malignancy have been analyzed to some extent by molecular genetic but not proteomics approaches. We screened sera of 94 melanoma patients for anti-melanoma reactivity and detected seropositivity in two-thirds of the patients with 2-6 antigens per case detected by 1D and an average of 2.3 per case by 2D Western blot analysis. For identification, antigen spots in Western blots were aligned with proteins in 2-DE and analyzed by mass spectrometry. 18 antigens were identified, 17 of which for the first time for melanoma. One of these antigens, galectin-3, has been related to various oncogenic processes including metastasis formation and invasiveness. Similarly, enolase has been found deregulated in different cancers. With at least 2 of 18 identified proteins implicated in oncogenic processes, the work confirms the potential of proteome-based antigen discovery to identify pathologically relevant proteins.

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Related in: MedlinePlus

Identification of melanoma-associated antigens detected by sera of melanoma patients.The protein spots that could be assigned to antigen spots in the Western blots (arrows here and in Figure 2) were excised from the gels and treated with trypsin. The resulting fragments were analyzed by mass spectrometry to identify the antigens. Eighteen different antigens were identified that were found in 46 different spots in the Western blots shown in Figure 2.
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pone-0005199-g003: Identification of melanoma-associated antigens detected by sera of melanoma patients.The protein spots that could be assigned to antigen spots in the Western blots (arrows here and in Figure 2) were excised from the gels and treated with trypsin. The resulting fragments were analyzed by mass spectrometry to identify the antigens. Eighteen different antigens were identified that were found in 46 different spots in the Western blots shown in Figure 2.

Mentions: Total protein extracts of M-NRT cells were separated with an isoelectric focusing pH range of 3–10 in the first and SDS-PAGE in the second dimension, blotted onto nitrocellulose membranes and probed with the sera of the patients as listed in Table 1. Serum dilutions were 1/200. The blots shown in Panels A through G were probed successively with 8 different patient sera each, blot H was probed with the sera of 9 healthy controls. The arrows indicated the antigens that could be assigned to protein spots in the silver-stained gel show with Figure 3. The numbering for the antigens is used throughout this report.


Proteome serological determination of tumor-associated antigens in melanoma.

Forgber M, Trefzer U, Sterry W, Walden P - PLoS ONE (2009)

Identification of melanoma-associated antigens detected by sera of melanoma patients.The protein spots that could be assigned to antigen spots in the Western blots (arrows here and in Figure 2) were excised from the gels and treated with trypsin. The resulting fragments were analyzed by mass spectrometry to identify the antigens. Eighteen different antigens were identified that were found in 46 different spots in the Western blots shown in Figure 2.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2667248&req=5

pone-0005199-g003: Identification of melanoma-associated antigens detected by sera of melanoma patients.The protein spots that could be assigned to antigen spots in the Western blots (arrows here and in Figure 2) were excised from the gels and treated with trypsin. The resulting fragments were analyzed by mass spectrometry to identify the antigens. Eighteen different antigens were identified that were found in 46 different spots in the Western blots shown in Figure 2.
Mentions: Total protein extracts of M-NRT cells were separated with an isoelectric focusing pH range of 3–10 in the first and SDS-PAGE in the second dimension, blotted onto nitrocellulose membranes and probed with the sera of the patients as listed in Table 1. Serum dilutions were 1/200. The blots shown in Panels A through G were probed successively with 8 different patient sera each, blot H was probed with the sera of 9 healthy controls. The arrows indicated the antigens that could be assigned to protein spots in the silver-stained gel show with Figure 3. The numbering for the antigens is used throughout this report.

Bottom Line: One of these antigens, galectin-3, has been related to various oncogenic processes including metastasis formation and invasiveness.Similarly, enolase has been found deregulated in different cancers.With at least 2 of 18 identified proteins implicated in oncogenic processes, the work confirms the potential of proteome-based antigen discovery to identify pathologically relevant proteins.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, Venerology and Allergy, Charité Universitätsmedizin Berlin, Humboldt University, Berlin, Germany.

ABSTRACT
Proteome serology may complement expression library-based approaches as strategy utilizing the patients' immune responses for the identification pathogenesis factors and potential targets for therapy and markers for diagnosis. Melanoma is a relatively immunogenic tumor and antigens recognized by melanoma-specific T cells have been extensively studied. The specificities of antibody responses to this malignancy have been analyzed to some extent by molecular genetic but not proteomics approaches. We screened sera of 94 melanoma patients for anti-melanoma reactivity and detected seropositivity in two-thirds of the patients with 2-6 antigens per case detected by 1D and an average of 2.3 per case by 2D Western blot analysis. For identification, antigen spots in Western blots were aligned with proteins in 2-DE and analyzed by mass spectrometry. 18 antigens were identified, 17 of which for the first time for melanoma. One of these antigens, galectin-3, has been related to various oncogenic processes including metastasis formation and invasiveness. Similarly, enolase has been found deregulated in different cancers. With at least 2 of 18 identified proteins implicated in oncogenic processes, the work confirms the potential of proteome-based antigen discovery to identify pathologically relevant proteins.

Show MeSH
Related in: MedlinePlus