Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample
Background Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both hum...
Ausführliche Beschreibung
Autor*in: |
Altara, Raffaele [verfasserIn] |
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E-Artikel |
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Sprache: |
Englisch |
Erschienen: |
2014 |
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Anmerkung: |
© Altara et al.; licensee BioMed Central Ltd. 2014 |
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Übergeordnetes Werk: |
Enthalten in: BMC biotechnology - London : BioMed Central, 2001, 14(2014), 1 vom: 15. Juli |
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Übergeordnetes Werk: |
volume:14 ; year:2014 ; number:1 ; day:15 ; month:07 |
Links: |
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DOI / URN: |
10.1186/1472-6750-14-63 |
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Katalog-ID: |
SPR028427076 |
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520 | |a Background Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. Results The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). Conclusions The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget. | ||
650 | 4 | |a Cytokines |7 (dpeaa)DE-He213 | |
650 | 4 | |a Chemokines |7 (dpeaa)DE-He213 | |
650 | 4 | |a Multiplex assay |7 (dpeaa)DE-He213 | |
650 | 4 | |a Planar assay |7 (dpeaa)DE-He213 | |
650 | 4 | |a Bead-based assay |7 (dpeaa)DE-He213 | |
650 | 4 | |a Inflammatory mediators |7 (dpeaa)DE-He213 | |
700 | 1 | |a Manca, Marco |4 aut | |
700 | 1 | |a Hessel, Marleen HM |4 aut | |
700 | 1 | |a Janssen, Ben J |4 aut | |
700 | 1 | |a Struijker-Boudier, Harry H A |4 aut | |
700 | 1 | |a Hermans, Rob JJ |4 aut | |
700 | 1 | |a Blankesteijn, W Matthijs |4 aut | |
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10.1186/1472-6750-14-63 doi (DE-627)SPR028427076 (SPR)1472-6750-14-63-e DE-627 ger DE-627 rakwb eng Altara, Raffaele verfasserin aut Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Altara et al.; licensee BioMed Central Ltd. 2014 Background Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. Results The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). Conclusions The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget. Cytokines (dpeaa)DE-He213 Chemokines (dpeaa)DE-He213 Multiplex assay (dpeaa)DE-He213 Planar assay (dpeaa)DE-He213 Bead-based assay (dpeaa)DE-He213 Inflammatory mediators (dpeaa)DE-He213 Manca, Marco aut Hessel, Marleen HM aut Janssen, Ben J aut Struijker-Boudier, Harry H A aut Hermans, Rob JJ aut Blankesteijn, W Matthijs aut Enthalten in BMC biotechnology London : BioMed Central, 2001 14(2014), 1 vom: 15. Juli (DE-627)332164837 (DE-600)2052746-9 1472-6750 nnns volume:14 year:2014 number:1 day:15 month:07 https://dx.doi.org/10.1186/1472-6750-14-63 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2119 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2014 1 15 07 |
spelling |
10.1186/1472-6750-14-63 doi (DE-627)SPR028427076 (SPR)1472-6750-14-63-e DE-627 ger DE-627 rakwb eng Altara, Raffaele verfasserin aut Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Altara et al.; licensee BioMed Central Ltd. 2014 Background Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. Results The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). Conclusions The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget. Cytokines (dpeaa)DE-He213 Chemokines (dpeaa)DE-He213 Multiplex assay (dpeaa)DE-He213 Planar assay (dpeaa)DE-He213 Bead-based assay (dpeaa)DE-He213 Inflammatory mediators (dpeaa)DE-He213 Manca, Marco aut Hessel, Marleen HM aut Janssen, Ben J aut Struijker-Boudier, Harry H A aut Hermans, Rob JJ aut Blankesteijn, W Matthijs aut Enthalten in BMC biotechnology London : BioMed Central, 2001 14(2014), 1 vom: 15. Juli (DE-627)332164837 (DE-600)2052746-9 1472-6750 nnns volume:14 year:2014 number:1 day:15 month:07 https://dx.doi.org/10.1186/1472-6750-14-63 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2119 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2014 1 15 07 |
allfields_unstemmed |
10.1186/1472-6750-14-63 doi (DE-627)SPR028427076 (SPR)1472-6750-14-63-e DE-627 ger DE-627 rakwb eng Altara, Raffaele verfasserin aut Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Altara et al.; licensee BioMed Central Ltd. 2014 Background Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. Results The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). Conclusions The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget. Cytokines (dpeaa)DE-He213 Chemokines (dpeaa)DE-He213 Multiplex assay (dpeaa)DE-He213 Planar assay (dpeaa)DE-He213 Bead-based assay (dpeaa)DE-He213 Inflammatory mediators (dpeaa)DE-He213 Manca, Marco aut Hessel, Marleen HM aut Janssen, Ben J aut Struijker-Boudier, Harry H A aut Hermans, Rob JJ aut Blankesteijn, W Matthijs aut Enthalten in BMC biotechnology London : BioMed Central, 2001 14(2014), 1 vom: 15. Juli (DE-627)332164837 (DE-600)2052746-9 1472-6750 nnns volume:14 year:2014 number:1 day:15 month:07 https://dx.doi.org/10.1186/1472-6750-14-63 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2119 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2014 1 15 07 |
allfieldsGer |
10.1186/1472-6750-14-63 doi (DE-627)SPR028427076 (SPR)1472-6750-14-63-e DE-627 ger DE-627 rakwb eng Altara, Raffaele verfasserin aut Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Altara et al.; licensee BioMed Central Ltd. 2014 Background Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. Results The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). Conclusions The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget. Cytokines (dpeaa)DE-He213 Chemokines (dpeaa)DE-He213 Multiplex assay (dpeaa)DE-He213 Planar assay (dpeaa)DE-He213 Bead-based assay (dpeaa)DE-He213 Inflammatory mediators (dpeaa)DE-He213 Manca, Marco aut Hessel, Marleen HM aut Janssen, Ben J aut Struijker-Boudier, Harry H A aut Hermans, Rob JJ aut Blankesteijn, W Matthijs aut Enthalten in BMC biotechnology London : BioMed Central, 2001 14(2014), 1 vom: 15. Juli (DE-627)332164837 (DE-600)2052746-9 1472-6750 nnns volume:14 year:2014 number:1 day:15 month:07 https://dx.doi.org/10.1186/1472-6750-14-63 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2119 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2014 1 15 07 |
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10.1186/1472-6750-14-63 doi (DE-627)SPR028427076 (SPR)1472-6750-14-63-e DE-627 ger DE-627 rakwb eng Altara, Raffaele verfasserin aut Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Altara et al.; licensee BioMed Central Ltd. 2014 Background Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. Results The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). Conclusions The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget. Cytokines (dpeaa)DE-He213 Chemokines (dpeaa)DE-He213 Multiplex assay (dpeaa)DE-He213 Planar assay (dpeaa)DE-He213 Bead-based assay (dpeaa)DE-He213 Inflammatory mediators (dpeaa)DE-He213 Manca, Marco aut Hessel, Marleen HM aut Janssen, Ben J aut Struijker-Boudier, Harry H A aut Hermans, Rob JJ aut Blankesteijn, W Matthijs aut Enthalten in BMC biotechnology London : BioMed Central, 2001 14(2014), 1 vom: 15. Juli (DE-627)332164837 (DE-600)2052746-9 1472-6750 nnns volume:14 year:2014 number:1 day:15 month:07 https://dx.doi.org/10.1186/1472-6750-14-63 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2119 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2014 1 15 07 |
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Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample Cytokines (dpeaa)DE-He213 Chemokines (dpeaa)DE-He213 Multiplex assay (dpeaa)DE-He213 Planar assay (dpeaa)DE-He213 Bead-based assay (dpeaa)DE-He213 Inflammatory mediators (dpeaa)DE-He213 |
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Altara, Raffaele Manca, Marco Hessel, Marleen HM Janssen, Ben J Struijker-Boudier, Harry H A Hermans, Rob JJ Blankesteijn, W Matthijs |
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improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample |
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Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample |
abstract |
Background Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. Results The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). Conclusions The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget. © Altara et al.; licensee BioMed Central Ltd. 2014 |
abstractGer |
Background Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. Results The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). Conclusions The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget. © Altara et al.; licensee BioMed Central Ltd. 2014 |
abstract_unstemmed |
Background Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. Results The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). Conclusions The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget. © Altara et al.; licensee BioMed Central Ltd. 2014 |
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Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample |
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Manca, Marco Hessel, Marleen HM Janssen, Ben J Struijker-Boudier, Harry H A Hermans, Rob JJ Blankesteijn, W Matthijs |
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