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  • Hoffmann, Holger (7)
  • Panne, Ulrich (4)
  • Schneider, Rudolf J. (3)
  • Baldofski, Stefanie (2)
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  • Esteves, V. I. (2)
  • Flemig, Sabine (2)
  • Hoffmann, Kristin (2)
  • Schneider, Rudolf (2)
  • Silva, C. P. (2)
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  • ELISA (9) (entfernen)

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LC-ELISA as a contribution to the validation of immunoassays (2019)
Schneider, Rudolf
Coupling ELISA and liquid chromatography yields a powerful too to detect cross-reacting substances, interfering matrix compounds and thus to validation of binding assays.
LC–ELISA as a contribution to the assessment of matrix effects with environmental water samples in an immunoassay for estrone (E1) (2018)
Hoffmann, Holger ; Knizia, Christian ; Kuhne, Maren ; Panne, Ulrich ; Schneider, Rudolf J.
Estrone (E1), a metabolite of the estrogenic hormones 17β-estradiol (β-E2) and 17α-estradiol (α-E2), is itself a potent estrogen which can have a significant impact on the hormonal balance. Due to ist high potential for adverse effects on human health and aquatic life even at pg/L to ng/L levels, its appearance in water should be monitored. E1 has also been considered a marker substance for the presence of other estrogens. This study presents a newly developed direct competitive enzymelinked immunosorbent assay (ELISA) for quantification of E1 in environmental water samples using new monoclonal antibodies. The quantification range of the ELISA is 0.15 μg/L to 8.7 μg/L E1, and the limit of detection is around 60 ng/L for not pre-concentrated water samples. A pre-concentration step after careful selection of suitable phases for SPE was developed, too. The influence of organic solvents and natural organic matter on the ELISA was assessed. The high selectivity of the monoclonal antibody was demonstrated by determining the cross-reactivity against 20 structurally related compounds. For the assessment of matrix effects, a concept (“LC–ELISA”) is thoroughly exploited, i.e., separating complex samples by HPLC into 0.3 min fractions and determination of the apparent E1 concentration. Furthermore, fractions with interferences for nontarget/suspected-target analysis can be assigned. A dilution approach was applied to distinguish between specific interferences (cross-reactants) and non-specific interferences (matrix effects). In the determination of 18 environmental samples, a good agreement of the E1 concentration in the respective fractions was obtained with mean recoveries of 103 % to 132 % comparing ELISA to LC–MS/MS.
Screening for cocaine on Euro banknotes by a highly sensitive enzyme immunoassay (2017)
Abdelshafi, Nahla A. ; Panne, Ulrich ; Schneider, Rudolf
This study focused on quantitative detection of cocaine on Euro banknotes in Germany. A sensitive direct competitive immunoassay was developed and optimized with a limit of detection (LOD) of 5.6 ng/L. Exhaustive cocaine extraction by solvent was tested using different methanol concentrations and buffered solutions. Crossreactivity studies were performed to determine the degree of interference of cocaine metabolites with the immunoassay. Sixty-five Euro banknotes obtained from different districts in Berlin were evaluated. A 100% contamination frequency with cocaine was detected. A comparison between the amount of cocaine extracted by cotton swabbing of one square centimeter of the banknote showed a good correlation for lower contamination levels. This assay showed high sensitivity of detecting pg of cocaine per 1 cm2 of one banknote by swabbing 1 cm2: 0, 14, and 21 pg/cm2. Moreover, three notes of different denominations revealed high cocaine concentration; 1.1 mg/note, and twice 55 μg/note.
Entwicklung und Anwendung eines Immunoassays (ELISA) zur Quantifizierung von Sulfamethoxazol in Abwasser und Oberflächengewässern (2016)
Hoffmann, Holger
N-Acetyl-SMX ist der Kreuzreaktand mit der bisher höchsten bekannten Kreuzreaktivität in diesem SMX-ELISA. Gemessene SMX-Konzentrationen in Klärwerkszuläufen waren im Bereich von 354-881 ng/L, in Abläufen von 221-421 ng/L und in Oberflächenwasser von 24-202 ng/L. Durch Verwendung einer Auftrennung der Probe in Fraktionen lassen sich störende Interferenzen beseitigen, aber die Probenmesszeit wird deutlich länger.
Structural considerations on the selectivity of an immunoassay for sulfamethoxazole (2016)
Hoffmann, Holger ; Baldofski, Stefanie ; Hoffmann, Kristin ; Flemig, Sabine ; Silva, C. P. ; Esteves, V. I. ; Emmerling, Franziska ; Panne, Ulrich ; Schneider, Rudolf J.
Sulfamethoxazol (SMX),a sulfonamide, is a widely used bacteriostatic antibiotic and therefore a promising marker for the entry of anthropogenic Pollution in the environment. SMX is frequently found in wastewater and surface water. This study presents the production of high affinity and selective polyclonal antibodies for SMX and the development and Evaluation of a direct competitive enzyme-linked immunosorbent assay(ELISA)for the quantification of SMX in environmental watersamples. The crystal structures of the cross-reacting compounds sulfamethizole, N4-acetyl-SMX andsuccinimidyl-SMX were determined by x-ray diffraction aiming to explain their high cross-reactivity. These crystal structures are described for the first time. The quantification range of the ELISA is 0.82–63 µg/L. To verify our results, the SMX concentration in 20 environmental samples,including wastewater and surfacewater,was determined by ELISA and tandem mass spectrometry(MS/MS).A good Agreement of the measured SMX concentrations was found with average recoveries of 97–113%for the results of ELISA compared to LC-MS/MS.
Behandlung von Klärwerkswasserproben zur Reduktion von Matrixeffekten bei der Quantifizierung von Sulfamethoxazol mittels Immunoassay (ELISA) (2016)
Hoffmann, Holger
N-Acetyl-SMX, der Kreuzreaktand mit der bisher höchsten bekannten Kreuzreaktivität in diesem SMX-ELISA, kann durch saure Behandlung entfernt werden. Durch Verdünnung der Probe kann die Überbestimmung reduziert werden, besonders für Proben mit hoher Interferenzbelastung (Matrix).
Entwicklung und Anwendung eines Immunoassays (ELISA) zur Quantifizierung von Sulfamethoxazol in Abwasser und Oberflächengewässern (2016)
Hoffmann, Holger
N-Acetyl-SMX ist der Kreuzreaktand mit der bisher höchsten bekannten Kreuzreaktivität in diesem SMX-ELISA. Gemessene SMX-Konzentrationen in Klärwerkszuläufen waren im Bereich von 354-881 ng/L, in Abläufen von 221-421 ng/L und in Oberflächenwasser von 24-202 ng/L. Durch Verwendung einer Auftrennung der Probe in Fraktionen lassen sich störende Interferenzen beseitigen, aber die Probenmesszeit wird deutlich länger.
Structural considerations on the selectivity of an immunoassay for sulfamethoxazole (2016)
Hoffmann, Holger ; Baldofski, Stefanie ; Hoffmann, Kristin ; Flemig, Sabine ; Silva, C. P. ; Esteves, V. I. ; Emmerling, Franziska ; Panne, Ulrich ; Schneider, Rudolf J.
Sulfamethoxazol (SMX),a sulfonamide, is a widely used bacteriostatic antibiotic and therefore a promising marker for the entry of anthropogenic Pollution in the environment. SMX is frequently found in wastewater and surface water. This study presents the production of high affinity and selective polyclonal antibodies for SMX and the development and Evaluation of a direct competitive enzyme-linked immunosorbent assay(ELISA)for the quantification of SMX in environmental watersamples. The crystal structures of the cross-reacting compounds sulfamethizole, N4-acetyl-SMX andsuccinimidyl-SMX were determined by x-ray diffraction aiming to explain their high cross-reactivity. These crystal structures are described for the first time. The quantification range of the ELISA is 0.82–63 µg/L. To verify our results, the SMX concentration in 20 environmental samples,including wastewater and surfacewater,was determined by ELISA and tandem mass spectrometry(MS/MS).A good Agreement of the measured SMX concentrations was found with average recoveries of 97–113%for the results of ELISA compared to LC-MS/MS.
Behandlung von Klärwerkswasserproben zur Reduktion von Matrixeffekten bei der Quantifizierung von Sulfamethoxazol mittels Immunoassay (ELISA) (2016)
Hoffmann, Holger
N-Acetyl-SMX, der Kreuzreaktand mit der bisher höchsten bekannten Kreuzreaktivität in diesem SMX-ELISA, kann durch saure Behandlung entfernt werden. Durch Verdünnung der Probe kann die Überbestimmung reduziert werden, besonders für Proben mit hoher Interferenzbelastung (Matrix).
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