TY - JOUR A1 - Mi, W. A1 - Josephs, R. D. A1 - Melanson, J. E. A1 - Dai, X. A1 - Wang, Y. A1 - Zhai, R. A1 - Chu, Z. A1 - Fang, X. A1 - Thibeault, M.-P. A1 - Stocks, B. B. A1 - Meija, J. A1 - Bedu, M. A1 - Martos, G. A1 - Westwood, S. A1 - Wielgosz, R. I. A1 - Liu, Q. A1 - Teo, T. L. A1 - Liu, H. A1 - Tan, Y. J. A1 - Öztuğ, M. A1 - Saban, E. A1 - Kinumi, T. A1 - Saikusa, K. A1 - Schneider, Rudolf A1 - Weller, Michael G. A1 - Konthur, Zoltán A1 - Jaeger, Carsten A1 - Quaglia, M. A1 - Mussell, C. A1 - Drinkwater, G. A1 - Giangrande, C. A1 - Vaneeckhoutte, H. A1 - Boeuf, A. A1 - Delatour, V. A1 - Lee, J. E. A1 - O'Connor, G. A1 - Ohlendorf, R. A1 - Henrion, A. A1 - Beltrão, P. J. A1 - Naressi Scapin, S. M. A1 - Sade, Y. B. T1 - PAWG Pilot Study on Quantification of SARS-CoV-2 Monoclonal Antibody - Part 1 N2 - Under the auspices of the Protein Analysis Working Group (PAWG) of the Comité Consultatif pour la Quantité de Matière (CCQM) a pilot study, CCQM-P216, was coordinated by the Chinese National Institute of Metrology (NIM), National Research Council of Canada (NRC) and the Bureau International des Poids et Mesures (BIPM). Eleven Metrology Institutes or Designated Institutes and the BIPM participated in the first phase of the pilot study (Part 1). The purpose of this pilot study was to develop measurement capabilities for larger proteins using a recombinant humanized IgG monoclonal antibody against Spike glycoprotein of SARS-CoV-2 (Anti-S IgG mAb) in solution. The first phase of the study was designed to employ established methods that had been previously studies by the CCQM Protein Analysis Working Group, involving the digestion of protein down to the peptide or amino acid level. The global coronavirus pandemic has also led to increased focus on antibody quantitation methods. IgG are among the immunoglobulins produced by the immune system to provide protection against SARS-CoV-2. Anti-SARS-CoV-2 IgG can therefore be detected in samples from affected patients. Antibody tests can show whether a person has been exposed to the SARS-CoV-2, and whether or not they potentially show lasting immunity to the disease. With the constant spread of the virus and the high pressure of re-opening economies, antibody testing plays a critical role in the fight against COVID-19 by helping healthcare professionals to identify individuals who have developed an immune response, either via vaccination or exposure to the virus. Many countries have launched large-scale antibody testing for COVID-19. The development of measurement standards for the antibody detection of SARS-CoV-2 is critically important to deal with the challenges of the COVID-19 pandemic. In this study, the SARS-CoV-2 monoclonal antibody is being used as a model system to build capacity in methods that can be used in antibody quantification. Amino acid reference values with corresponding expanded uncertainty of 36.10 ± 1.55 mg/kg, 38.75 ± 1.45 mg/kg, 18.46 ± 0.78 mg/kg, 16.20 ± 0.67 mg/kg and 30.61 ± 1.30 mg/kg have been established for leucine, valine, phenylalanine, isoleucine and proline, respectively. Agreement between nearly all laboratories was achieved for the amino acid analysis within 2 to 2.5 %, with one participant achieving markedly higher results due to a technical issue found in their procedure; this result was thus excluded from the reference value calculations. The relatively good agreement within a laboratory between different amino acids was not dissimilar to previous results for peptides or small proteins, indicating that factors such as hydrolysis conditions and calibration procedures could be the largest sources of variability. Peptide reference values with corresponding expanded uncertainty of 4.99 ± 0.28 mg/kg and 6.83 ± 0.65 mg/kg have been established for ALPAPIEK and GPSVFPLAPSSK, respectively. Not surprisingly due to prior knowledge from previous studies on peptide quantitation, agreement between laboratories for the peptide-based analysis was slightly poorer at 3 to 5 %, with one laboratory's result excluded for the peptide GPSVFPLAPSSK. Again, this level of agreement was not significantly poorer than that achieved in previous studies with smaller or less complex proteins. To reach the main text of this paper, click on Final Report. KW - Antibody quantification KW - Amino acid analysis KW - Peptide analysis KW - Round robin test PY - 2021 DO - https://doi.org/10.1088/0026-1394/59/1a/08001 VL - 59 IS - 1A SP - 08001 AN - OPUS4-54972 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Pereira, P.M.R. A1 - Carvalho, José Joao A1 - Silva, S. A1 - Cavaleiro, J.A.S. A1 - Schneider, Rudolf A1 - Fernandes, R. A1 - Tomé, J.P.C. T1 - Porphyrin conjugated with serum albumins and monoclonal antibodies boosts efficiency in targeted destruction of human bladder cancer cells N2 - The synthesis of a novel PS conjugated with bovine and human serum albumin (BSA and HSA) and a monoclonal antibody anti-CD104 is reported, as well as their biological potential against the human bladder cancer cell line UM-UC-3. No photodynamic effect was detected when the non-conjugated porphyrin was used. Yet, when it was coupled covalently with the mAb anti-CD104, BSA and HSA, the resulting photosensitizer conjugates demonstrated high efficacy in destroying the cancer cells, the mAb anti-CD104 efficacy overruling the albumins. KW - Rinderserumalbumin KW - BSA KW - Monoklonale Antikörper KW - Blasenkrebs KW - Konjugate KW - Conjugates KW - Therapeutische Antikörper KW - CD104 PY - 2014 DO - https://doi.org/10.1039/c3ob42082e SN - 1477-0520 SN - 1477-0539 VL - 12 IS - 11 SP - 1804 EP - 1811 PB - RSC CY - Cambridge AN - OPUS4-30451 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Scala-Benuzzi, M. L. A1 - Soler-Illia, G. J. A. A. A1 - Rabia, J. A1 - Battaglini, F. A1 - Schneider, Rudolf A1 - Pereira, S. V. A1 - Messina, G. A. T1 - Immunosensor based on porous gold and reduced graphene platform for the determination of EE2 by electrochemical impedance spectroscopy N2 - In this work, we report an electrochemical immunosensor to detect ethinylestradiol in water samples, using electrochemical impedance spectroscopy (EIS) as a detection technique. For the development of this immunosensor, the direct modification of the working electrode of a screen-printed carbon electrode was carried out. First, to reduce the resistance of the electrode, electroreduced graphene was incorporated on the surface. Second, a porous gold structure was electrodeposited on reduced graphene by electrodeposition and the dynamic hydrogen bubble template assisted method. Thus, a marked increase in surface area was obtained for anti-EE2 antibodies immobilization. Subsequently, the specific anti-EE2 antibodies were covalently immobilized using α-lipoic acid for attaching them to the gold surface. The electrode modified with the antibodies was incubated for 30 min in the samples containing EE2, producing the specific Antigen antibody binding. As the charge transfer resistance of a redox probe in the electrode surface is governed by the surface blocking effects, the charge transfer resistance was related to the amount of EE2 captured to realize a quantitative determination. For this, the EIS measurements were performed in a 4 mM [Fe(CN)6]4−/3− solution in 0.1 M KCl. The obtained Nyquist diagrams were adjusted using the Randles circuit as an equivalent circuit to obtain the corresponding resistances. The developed methodology showed good selectivity, precision, and sensitivity; although the LOD obtained was higher than those presented in other published articles, it turned out to be an alternative that allows the determination of ethinylestradiol using a simple disposable electrode. KW - Ethinylestradiol KW - Biosensor KW - Elektrochemisch KW - Impedanz PY - 2021 DO - https://doi.org/10.1016/j.jelechem.2021.115604 SN - 1572-6657 VL - 897 SP - 115604 EP - 115611 PB - Elsevier B.V. CY - Amsterdam AN - OPUS4-54048 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schneider, Rudolf A1 - Korsak, B. A1 - Almeida, G. M. A1 - Rocha, S. A1 - Pereira, C. A1 - Mendes, N. A1 - Osorio, H. A1 - Pereira, P. M. A1 - Rodrigues, J. M. M. A1 - Sarmento, B. A1 - Tome, J. P. C. A1 - Oliveira, C. T1 - Porphyrin modified trastuzumab improves efficacy of HER2 targeted photodynamic therapy of gastric cancer N2 - Gastric cancer (GC) is the 3rd deadliest cancer worldwide, due to limited treatment options and late diagnosis. Human epidermal growth factor receptor-2 (HER2) is overexpressed in similar to 20% of GC cases and anti-HER2 antibody trastuzumab in combination with conventional chemotherapy, is recognized as standard therapy for HER2-positive metastatic GC. This strategy improves GC patients' survival by 2-3 months, however its optimal results in breast cancer indicate that GC survival may be improved. A new photoimmunoconjugate was developed by conjugating a porphyrin with trastuzumab (Trast: Porph) for targeted photodynamic therapy in HER2-positive GC. Using mass spectrometry analysis, the lysine residues in the trastuzumab structure most prone for porphyrin conjugation were mapped. The in vitro data demonstrates that Trast: Porph specifically binds to HER2-positive cells, accumulates intracellularly, co-localizes with lysosomal marker LAMP1, and induces massive HER2-positive cell death upon cellular irradiation. The high selectivity and cytotoxicity of Trast: Porph based photoimmunotherapy is confirmed in vivo in comparison with trastuzumab alone, using nude mice xenografted with a HER2-positive GC cell line. In the setting of human disease, these data suggest that repetitive cycles of Trast: Porph photoimmunotherapy may be used as an improved treatment strategy in HER2-positive GC patients. KW - Photoimmunotherapy KW - Photoimmunoconjugate KW - Gastric cancer PY - 2017 DO - https://doi.org/10.1002/ijc.30844 SN - 0020-7136 VL - 141 IS - 7 SP - 1478 EP - 1489 PB - International Journal of Cancer AN - OPUS4-43315 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Scala-Benuzzi, M. L. A1 - Takara, E. A. A1 - Alderete, M. A1 - Soler-Illia, G. J. A. A. A1 - Schneider, Rudolf A1 - Raba, J. A1 - Messina, G. A. T1 - Ethinylestradiol quantification in drinking water sources using a fluorescent paper based immunosensor N2 - In this work we report a novel paper-based analytical device read-out via LED-induced fluorescence detection (FPAD) for the quantification of the emerging pollutant ethinylestradiol (EE2) in river water samples. The PAD was used as a reaction platform for a competitive enzyme immunoassay. For the PAD development, microzones of filter paper, printed by a wax printing method, were modified with amino-functionalized SBA-15 and subsequently, anti-EE2 specific antibodies were covalently immobilized. The determination of EE2 in water was carried out by adding a fixed concentration of EE2 conjugated with the enzyme horseradish peroxidase (HRP) to samples and standards. Then, the FPAD were added and incubated for 10 min. Finally, the detection was performed by the reaction of 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) whose oxidation is catalyzed by HRP in the presence of H2O2, obtaining the highly fluorescent resorufin (R). Resorufin was detected by LED excitation at 550 nm, observing emission at 585 nm. The EE2 concentration in the samples was inversely proportional to the relative fluorescence obtained from the enzymatic reaction products. The FPAD assay showed a detection Limit (LOD) of 0.05 ng L−1 and coefficients of variation (CV) below 4.5% within-assay and below 6.5% between-assay, respectively. The results obtained show the potential suitability of our FPAD for the selective and sensitive quantification of EE2 in river water samples. In addition, it has the PADs advantages of being disposable, easy to apply and inexpensive. KW - Immunoassay KW - Arzneimittel KW - Antikörper KW - LED-induzierte Fluoreszenz KW - Paper based devices KW - Mesoporöses Silica KW - Partikel PY - 2018 DO - https://doi.org/10.1016/j.microc.2018.05.038 SN - 0026-265X VL - 141 SP - 287 EP - 293 PB - Elsevier B.V. CY - Amsterdam, NL AN - OPUS4-45269 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Sin, DWM. A1 - Wong, YC. A1 - Lehmann, Andreas A1 - Schneider, Rudolf A1 - Kakoulides, E. A1 - Lin, TT. A1 - Liu, QD. A1 - Cabillic, J. A1 - Lardy-fontan, S. A1 - Nammoonnoy, J. T1 - CCQM-K126: low polarity organic in water: carbamazepine in surface water N2 - This study aimed to assess the measurement capabilities of participating National Metrology Institutes/ Designated Institutes (NMIs/DIs) and expert laboratories in determining of low-polarity organics in surface water. This comparison was organized by Government Laboratory, Hong Kong (GLHK). At the CCQM Organic Analysis Working Group (OAWG) Meeting held in November 2012 in Hong Kong, GLHK initially proposed a CCQM key comparison and a parallel pilot study programme on pharmaceuticals in surface water. Further discussion at the CCQM Meeting held in April 2014 in Paris, the OAWG approved a CCQM Track C comparison (CCQM-K126) on low polarity pharmaceuticals in surface water. In the meeting, the programme was supported by more than six National Metrology Institutes/ Designated Institutes (NMIs/ DIs). CCQM-K126 officially commenced in July 2014 and had registration from nine NMIs/DIs. Participants were provided two bottles (40 mL each) of surface water and were requested to determine the mass fraction of spiked carbamazepinein in surface water. The coordinator received nine sets of results from eight NMIs/DIs in February 2015. Apart from one using an immunoassay technique, all participants applied isotope dilution liquid chromatography-tandem mass spectrometry (ID-LCMS/MS) technique as their determination technique. KW - CCQM-K126 KW - ELISA KW - Mass spectrometry PY - 2017 DO - https://doi.org/10.1088/0026-1394/54/1A/08030 SN - 0026-1394 SN - 1681-7575 VL - 54 IS - Suppl. S SP - Article 08030, 1 EP - 56 PB - IOP Publ. CY - Bristol AN - OPUS4-44831 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Scala-Benuzzi, M. L. A1 - Raba, J. A1 - Soler-Illia, G. J. A. A. A1 - Schneider, Rudolf A1 - Messina, G. A. T1 - Novel electrochemical paper-based immunocapture assay for the quantitative determination of ethinylestradiol in water samples N2 - We report a novel and innovative electrochemical paper-based immunocapture assay (EPIA) to address the need for ultrasensitive detection of emerging pollutants without regulatory status and whose effects on environment and human health are not completely yet understood. In particular, we present the application of this system toward highly sensitive detection of the emerging pollutant ethinyl estradiol (EE2). The EPIA approach is based on the use of paper microzones modified with silica nanoparticles (SNs) and anti-EE2 specific antibodies for capture and preconcentration of EE2 from river water samples. After the preconcentration procedure, the paper microzones are placed onto a screen-printed carbon electrode modified with electrochemically reduced graphene (RG). The bound EE2 is subsequently desorbed adding a diluted solution of sulfuric acid on the paper microzones. Finally, recovered EE2 is electrochemically detected by OSWV. The proposed novel methodology showed an appropriate LOD and linear range for the quantification of EE2 for water samples with different origins. The nonsophisticated equipment required, the adequate recovery values obtained (from 97% to 104%, with a RSD less than 4.9%), and the appropriate LOD and linear range value (0.1 ng L−1 and 0.5−120 ng L−1, respectively) achieved by our immunocapture sensor present significant analytical figures of merit, particularly when the routine quantification of EE2 is considered. In addition, our System was based on electrochemical paper-based technology, which allows obtainment of portable, easy-to-use, inexpensive, and disposable devices. The EPIA can also serve as a general-purpose immunoassay platform applicable to quantitation of other drugs and emerging pollutants in environmental samples. KW - Immunoassay KW - EE2 KW - Estrogens KW - Screen-printed electrode KW - Square-wave voltammetry PY - 2018 DO - https://doi.org/10.1021/acs.analchem.8b00028 SN - 0003-2700 SN - 1520-6882 VL - 90 IS - 6 SP - 4104 EP - 4111 PB - American Chemical Society CY - Washington, D.C., USA AN - OPUS4-44690 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Wang, L. A1 - Stebbings, R. A1 - Gaigalas, A.K. A1 - Sutherland, J. A1 - Kammel, M. A1 - John, M. A1 - Roemer, B. A1 - Kuhne, Maren A1 - Schneider, Rudolf A1 - Braun, M. A1 - Engel, A. A1 - Dikshit, D. A1 - Abbasi, F. A1 - Marti, G.E. A1 - Sassi, M. A1 - Revel, L. A1 - Kim, S.K. A1 - Baradez, M.-O. A1 - Lekishvili, T. A1 - Marshall, D. A1 - Whitby, L. A1 - Jing, W. A1 - Ost, V. A1 - Vonsky, M. A1 - Neukammer, J. T1 - Quantification of cells with specific phenotypes II: Determination of CD4 expression level on reconstituted lyophilized human PBMC labelled with anti-CD4 FITC antibody N2 - This report focuses on the characterization of CD4 expression level in terms of equivalent number of reference fluorophores (ERF). Twelve different flow cytometer platforms across sixteen laboratories were utilized in this study. As a first step the participants were asked to calibrate the fluorescein isothiocyanate (FITC) channel of each flow cytometer using commercially available calibration standard consisting of five populations of microspheres. Each population had an assigned value of equivalent fluorescein fluorophores (EFF denotes a special case of the generic term ERF with FITC as the reference fluorophore). The EFF values were assigned at the National Institute of Standards and Technology (NIST). A surface-labelled lyophilized cell preparation was provided by the National Institute of Biological Standards and Control (NIBSC), using human peripheral blood mononuclear cells (PBMC) pre-labeled with a FITC conjugated anti-CD4 monoclonal antibody. Three PBMC sample vials, provided to each participant, were used for the CD4 expression analysis. The PBMC are purported to have a fixed number of surface CD4 receptors. On the basis of the microsphere calibration, the EFF value of the PBMC samples was measured to characterize the population average CD4 expression level of the PBMC preparations. Both the results of data analysis performed by each participant and the results of centralized analysis of all participants' raw data are reported. Centralized analysis gave a mean EFF value of 22,300 and an uncertainty of 750, corresponding to 3.3% (level of confidence 68%) of the mean EFF value. The next step will entail the measurement of the ERF values of the lyophilized PBMC stained with labels for other fluorescence channels. The ultimate goal is to show that lyophilized PBMC is a suitable biological reference cell material for multicolor flow cytometry and that it can be used to present multicolor flow cytometry measurements in terms of ABC (antibodies bound per cell) units. KW - Surface labelled lyophilized PBMC KW - CD4 expression level KW - FITC KW - Equivalent fluorescein fluorophore (EFF) KW - Quantitative flow cytometry KW - Calibration KW - Standard measurement procedure KW - Measurement uncertainty KW - Reference cell material PY - 2015 DO - https://doi.org/10.1002/cyto.a.22634 SN - 0196-4763 SN - 1552-4922 SN - 1552-4930 VL - 87 IS - 3 SP - 254 EP - 261 PB - Wiley-Liss CY - Hoboken, NJ AN - OPUS4-32981 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Stebbings, R. A1 - Wang, L. A1 - Sutherland, J. A1 - Kammel, M. A1 - Gaigalas, A.K. A1 - John, M. A1 - Roemer, B. A1 - Kuhne, Maren A1 - Schneider, Rudolf A1 - Braun, M. A1 - Engel, A. A1 - Dikshit, D.K. A1 - Abbasi, F. A1 - Marti, G.E. A1 - Sassi, M.P. A1 - Revel, L. A1 - Kim, S.-K. A1 - Baradez, M.-O. A1 - Lekishvili, T. A1 - Marshall, D. A1 - Whitby, L. A1 - Jing, W. A1 - Ost, V. A1 - Vonsky, M. A1 - Neukammer, J. T1 - Quantification of cells with specific phenotypes I: Determination of CD4+ cell count per microliter in reconstituted lyophilized human PBMC prelabeled with anti-CD4 FICT antibody N2 - A surface-labeled lyophilized lymphocyte (sLL) preparation has been developed using human peripheral blood mononuclear cells prelabeled with a fluorescein isothiocyanate conjugated anti-CD4 monoclonal antibody. The sLL preparation is intended to be used as a reference material for CD4+ cell counting including the development of higher order reference measurement procedures and has been evaluated in the pilot study CCQM-P102. This study was conducted across 16 laboratories from eight countries to assess the ability of participants to quantify the CD4+ cell count of this reference material and to document cross-laboratory variability plus associated measurement uncertainties. Twelve different flow cytometer platforms were evaluated using a standard protocol that included calibration beads used to obtain quantitative measurements of CD4+ T cell counts. There was good overall cross-platform and counting method agreement with a grand mean of the laboratory calculated means of (301.7 ± 4.9) µL-1 CD4+ cells. Excluding outliers, greater than 90% of participant data agreed within ±15%. A major contribution to variation of sLL CD4+ cell counts was tube to tube variation of the calibration beads, amounting to an uncertainty of 3.6%. Variation due to preparative steps equated to an uncertainty of 2.6%. There was no reduction in variability when data files were centrally reanalyzed. Remaining variation was attributed to instrument specific differences. CD4+ cell counts obtained in CCQM-P102 are in excellent agreement and show the robustness of both the measurements and the data analysis and hence the suitability of sLL as a reference material for interlaboratory comparisons and external quality assessment. KW - CD4+ cell counting KW - Relative concentration measurement KW - Lyophilized cells KW - Flow cytometry KW - Standard measurement procedure KW - Measurement of uncertainty KW - Human immunodeficiency virus-1 KW - Acquired immunodeficiency syndrome KW - Reference material PY - 2015 DO - https://doi.org/10.1002/cyto.a.22614 SN - 0196-4763 SN - 1552-4922 SN - 1552-4930 VL - 87 IS - 3 SP - 244 EP - 253 PB - Wiley-Liss CY - Hoboken, NJ AN - OPUS4-32847 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Carvalho, J.J.D. A1 - Bahlmann, Arnold A1 - Schneider, Rudolf T1 - ELISA goes Berlin - Bestimmung von Carbamazepin, Cetrizin und Coffein in Berliner Gewässern N2 - Sensitive und selektive Immunoassays unterstützen Untersuchungen im Vollzug der EU Wasserrahmenrichtlinie. Sie erlauben, eine hohe Anzahl von Proben ohne Anreicherungsschritte zu untersuchen und ermöglichen daher den Aufbau eines zeitlich und räumlich engmaschigen Messnetzes. Mit Hilfe von ELISA (enzymelinked immunosorbent assay) wurde ein Gewässerscreening in Berlin durchgeführt. KW - ELISA KW - Koffein KW - Immunoassay KW - Oberflächenwasser KW - Wasseranalytik PY - 2012 SN - 0016-3538 VL - 4 SP - 278 EP - 281 PB - GIT-Verlag CY - Darmstadt AN - OPUS4-27518 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Gärtner, Stefanie A1 - Carvalho, J.J. A1 - Emmerling, Franziska A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Polyclonal murine and rabbit antibodies for the bile acid isolithocholic acid N2 - Bile acids are relevant markers for clinical research. This study reports the production of antibodies for isolithocholic acid, the isomer of the extensively studied lithocholic acid. The IgG titer and affinity maturation were monitored during the immunizations of three mice and two rabbits. In both animal models, polyclonal antibodies with a high selectivity and affinity were produced. The development of a direct competitive ELISA with a test midpoint of 0.69 ± 0.05 µg/L and a measurement range from 0.09–15 µg/L is reported. Additionally, the crystal structure of isolithocholic acid is described for the first time. KW - ELISA KW - Crystal structure KW - Competitive assay KW - Immunization monitoring KW - 3ß-hydroxy-5ß-cholanic acid PY - 2015 DO - https://doi.org/10.1080/15321819.2014.924419 SN - 1532-1819 SN - 1532-4230 VL - 36 IS - 3 SP - 233 EP - 252 PB - Taylor & Francis CY - Philadelphia, Pa. AN - OPUS4-31836 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Freitas, R. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Velez, C. A1 - Moreira, A. A1 - Schneider, Rudolf A1 - Esteves, V.I. A1 - Wrona, F. J. A1 - Soares, A.M.V.M. A1 - Figueira, E. T1 - How life history influences the responses of the clam Scrobicularia plana to the combined impacts of carbamazepine and pH decrease N2 - In the present study, the bivalve Scrobicularia plana, collected from two contrasting areas (pristine location and mercury contaminated area), was selected to assess the biochemical alterations imposed by pH decrease, carbamazepine (an antiepileptic) and the combined effect of both stressors. The effects on oxidative stress related biomarkers after 96 h exposure revealed that pH decrease and carbamazepine induced alterations on clams, with greater impacts on individuals from the contaminated area which presented higher mortality, higher lipid peroxidation and higher glutathione S-transferase activity. These results emphasize the risk of extrapolating results from one area to another, since the same species inhabiting different areas may be affected differently when exposed to the same stressors. Furthermore, the results obtained showed that, when combined, the impact of pH decrease and carbamazepine was lower than each stressor acting alone, which could be related to the defence mechanism of valves closure when bivalves are under higher stressful conditions. KW - Ocean acidification KW - Biomarkers KW - Oxidative stress KW - Bivalves KW - Pharmaceutical drugs PY - 2015 DO - https://doi.org/10.1016/j.envpol.2015.03.023 SN - 0269-7491 SN - 0013-9327 SN - 1873-6424 VL - 202 SP - 205 EP - 214 PB - Elsevier CY - New York, NY [u.a.] AN - OPUS4-33818 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Ecke, Alexander A1 - Westphalen, Tanja A1 - Hornung, J. A1 - Voetz, M. A1 - Schneider, Rudolf T1 - A rapid magnetic bead-based immunoassay for sensitive determination of diclofenac N2 - Increasing contamination of environmental waters with pharmaceuticals represents an emerging threat for the drinking water quality and safety. In this regard, fast and reliable analytical methods are required to allow quick countermeasures in case of contamination. Here, we report the development of a magnetic bead-based immunoassay (MBBA) for the fast and cost-effective determination of the analgesic diclofenac (DCF) in water samples, based on diclofenac-coupled magnetic beads and a robust monoclonal anti-DCF antibody. A novel synthetic strategy for preparation of the beads resulted in an assay that enabled for the determination of diclofenac with a significantly lower limit of detection (400 ng/L) than the respective enzyme-linked immunosorbent assay (ELISA). With shorter incubation times and only one manual washing step required, the assay demands for remarkably shorter time to result (< 45 min) and less equipment than ELISA. Evaluation of assay precision and accuracy with a series of spiked water samples yielded results with low to moderate intra- and inter-assay variations and in good agreement with LC–MS/MS reference analysis. The assay principle can be transferred to other, e.g., microfluidic, formats, as well as applied to other analytes and may replace ELISA as the standard immunochemical method. KW - Immunoassay KW - Magnetic beads KW - Diclofenac KW - Water analysis KW - LC-MS/MS PY - 2021 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-542346 DO - https://doi.org/10.1007/s00216-021-03778-7 SN - 1618-2650 VL - 414 SP - 1563 EP - 1573 PB - Springer CY - Heidelberg AN - OPUS4-54234 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Gregório, Bruno J.R. A1 - Ramos, Inês I. A1 - Marques, Sara S. A1 - Barreiros, Luísa A1 - Magalhães, Luís M. A1 - Schneider, Rudolf A1 - Segundo, Marcela A. T1 - Microcarrier-based fluorescent yeast estrogen screen assay for fast determination of endocrine disrupting compounds N2 - The presence of endocrine-disrupting compounds (EDCs) in water poses a significant threat to human and animal health, as recognized by regulatory agencies throughout the world. The Yeast Estrogen Screen (YES) assay is an excellent method to evaluate the presence of these compounds in water due to its simplicity and capacity to assess the bioaccessible forms/fractions of these compounds. In the presence of a compound with estrogenic activity, Saccharomyces cerevisiae cells, containing a lacZ reporter gene encoding the enzyme β-galactosidase, are induced, the enzyme is synthesised, and released to the extracellular medium. In this work, a YES-based approach encompassing the use of a lacZ reporter gene modified strain of S. cerevisiae, microcarriers as solid support, and a fluorescent substrate, fluorescein di-β-D-galactopyranoside, is proposed, allowing for the assessment of EDCs’ presence after only 2 h of incubation. The proposed method provided an EC50 of 0.17 ± 0.03 nM and an LLOQ of 0.03 nM, expressed as 17β-estradiol. The assessment of different EDCs provided EC50 values between 0.16 and 1.2 × 103 nM. After application to wastewaters, similar results were obtained for EDCs screening, much faster, compared to the conventional 45 h spectrophotometric procedure using a commercial kit, showing potential for onsite high-throughput screening of environmental contamination. KW - Biosensoren KW - YES assay KW - Endokrine Disruptoren PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-602315 DO - https://doi.org/10.1016/j.talanta.2024.125665 VL - 271 SP - 1 EP - 7 PB - Elsevier B.V. CY - Amsterdam AN - OPUS4-60231 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Pieres, A. A1 - Almeida, Ângela A1 - Correia, J. A1 - Calisto, V. A1 - Schneider, Rudolf A1 - Esteves, V. I. A1 - Soares, A. M. V. M. A1 - Figueira, E. A1 - Freitas, R. T1 - Long-term exposure to caffeine and carbamazepine: Impacts on the regenerative capacity of the polychaete Diopatra neapolitana N2 - The toxicity induced in non-target organisms by pharmaceutical drugs has been the focus of several studies. In the aquatic environment, most of the studies have been devoted to fish and bivalves, while little is known on the impacts induced in polychaetes. The present study evaluated the impacts of carbamazepine and caffeine on the regenerative capacity of Diopatra neapolitana, a polychaete species with high ecological and economic relevance. Under laboratory controlled conditions polychaetes were exposed, during 28 days, to carbamazepine (Ctl-0.0; 0.3; 3.0; 6.0; 9.0 mg/L) and caffeine (Ctl-0.0; 0.5; 3.0; 18.0 mg/L). During the experiment, at days 11, 18, 25, 32, 39 and 46 after amputation, for each specimen, the percentage of the body width regenerated was determined and the number of new segments was counted. The regenerative capacity was assessed considering the number of days needed to achieve full regeneration and the total number of new segments. The obtained results revealed that with the increase of drugs concentrations organisms regenerated less new segments and took longer to completely regenerate. KW - Diopatra neapolitana KW - Pollution KW - Pharmaceutical drugs KW - Regenerative capacity PY - 2016 DO - https://doi.org/10.1016/j.chemosphere.2015.12.035 SN - 0045-6535 VL - 146 SP - 565 EP - 573 PB - Elsevier AN - OPUS4-38497 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Freitas, R. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Velez, C. A1 - Moreira, A. A1 - Schneider, Rudolf A1 - Esteves, V. I. A1 - Wrona, F. J. A1 - Figueira, E. A1 - Soares, A. M. V. M. T1 - The impacts of pharmaceutical drugs under ocean acidification: Newdata on single and combined long-term effects of carbamazepine on Scrobicularia plana N2 - Ocean acidification and increasing discharges of pharmaceutical contaminants into aquatic systems are among key and/or emerging drivers of environmental change affecting marine ecosystems. A growing body of evidence demonstrates that ocean acidification can have direct and indirect impacts on marine organisms although combined effects with other stressors, namely with pharmaceuticals, have received very little attention to date. The present study aimed to evaluate the impacts of the pharmaceutical drug Carbamazepine and pH 7.1, acting alone and in combination, on the clamScrobicularia plana. For this, a long-termexposure (28 days)was conducted and a set of oxidative stress markers was investigated. The results obtained showed that S. plana was able to develop mechanisms to prevent oxidative damage when under low pH for a long period, presenting higher survival when exposed to this stressor compared to CBZ or the combination of CBZ with pH 7.1. Furthermore, the toxicity of CBZ on S. plana was synergistically increased under ocean acidification conditions (CBZ + pH 7.1): specimens survival was reduced and oxidative stress was enhanced when compared to single exposures. These findings add to the growing body of evidence that ocean acidification will act to increase the toxicity of CBZ to marine organisms,which has clear implications for coastal benthic ecosystems suffering chronic pollution from pharmaceutical drugs. KW - Ocean acidification KW - Pharmaceuticals KW - Biomarkers KW - Oxidative stress KW - Clams KW - Long-term exposures PY - 2016 DO - https://doi.org/10.1016/j.scitotenv.2015.09.138 VL - 541 SP - 977 EP - 985 PB - Elsevier B.V. AN - OPUS4-38502 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Fernandez, L. A1 - Esteves, V. I. A1 - Cunha, A. A1 - Schneider, Rudolf A1 - Tome, J. P. C. T1 - Photodegradation of organic pollutants in water by immobilized porphyrins and phthalocyanines N2 - New methods for water treatment are required as a result from an increasing awareness in the reduction of the pollution impact in the environment. In the perspective of the photo-oxidation of organic pollutants present in water, the principal incentive for the preparation of heterogeneous photocatalysts is their easy recovery from the reaction mixture, which allows their reuse in successive runs, minimizing the loss of their original photocatalytic properties. Different types of supports can be used in the immobilization of photoactive species, such as porphyrins (Pors) and phthalocyanines (Pcs). This mini-review will consider the different methodologies for the immobilization of Pors and Pcs and their photocatalytic performance in the photodegradation of organic pollutants in water, addressing also their recycling ability in successive water treatments. KW - Porphyrins KW - Phthalocyanines KW - Water treatment KW - Organic pollutants KW - Advanced oxidation processes KW - Heterogeneous photocatalysis KW - TiO2 KW - Microporous KW - Nanoparticles PY - 2016 DO - https://doi.org/10.1142/S108842461630007X VL - 2016 IS - 20 SP - 150 EP - 166 PB - World Scientific Publishing AN - OPUS4-38503 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Pires, A. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Schneider, Rudolf A1 - Esteves, V. I. A1 - Wrona, F. J. A1 - Soares, A. M. V. M. A1 - Figueira, E. A1 - Freitas, R. T1 - Long-term exposure of polychaetes to caffeine: Biochemical alterations induced in Diopatra neapolitana and Arenicola marina N2 - In the last decade studies have reported the presence of several pharmaceutical drugs in aquatic environments worldwide and an increasing effort has been done to understand the impacts induced on wildlife. Among the most abundant drugs in the environment is caffeine, which has been reported as an effective chemical anthropogenic marker. However, as for the majority of pharmaceuticals, scarce information is available on the adverse effects of caffeine on marine benthic organisms, namely polychaetes which are the most abundant group of organisms in several aquatic ecossystems. Thus, the present study aimed to evaluate the biochemical alterations induced by environmentally relevant concentrations of caffeine on the polychaete species Diopatra neapolitana and Arenicola marina. The results obtained demonstrated that after 28 days exposure oxidative stress was induced in both species, especially noticed in A. marina, resulting from the incapacity of antioxidant and biotransformation enzymes to prevent cells from lipid peroxidation. The present study further revealed that D. neapolitana used glycogen and proteins as energy to develop defense mechanisms while in A. marina these reserves were maintained independently on the exposure concentration, reinforcing the low capacity of this species to fight against oxidative stress. KW - Invertebrates KW - Pharmaceuticals KW - Oxidative stress biomarkers KW - Energy reserves PY - 2016 DO - https://doi.org/10.1016/j.envpol.2016.04.031 VL - 2016 IS - 214 SP - 456 EP - 463 PB - Elsevier Ltd. AN - OPUS4-38505 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Cruz, D. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Esteves, V. I. A1 - Schneider, Rudolf A1 - Wrona, F. J. A1 - Soares, A. M. V. M. A1 - Figueira, E. A1 - Freitas, R. T1 - Caffeine impacts in the clam Ruditapes philippinarum: Alterations on energy reserves, metabolic activity and oxidative stress biomarkers N2 - Caffeine is known to be one of the most consumed psychoactive drugs. For this reason, caffeine is continuously released into the environment with potential impacts on inhabiting organisms. The current study evaluated the biochemical alterations induced in the clam species Ruditapes philippinarum after exposure for 28 days to caffeine (0.5, 3.0 and 18.0 mg/L). The results obtained showed that, with the increasing caffeine concentrations, an increase in clams defense mechanisms (such as antioxidant and biotransformation enzymes activity) was induced which was accompanied by an increase in protein content. Nevertheless, although an increase on defense mechanisms was observed, clams were not able to prevent cells from lipid peroxidation that increased with the increase of caffeine concentration. Furthermore, with the increase of exposure concentrations, clams increased their metabolic activity (measured by electron transport activity), reducing their energy reserves (glycogen content), to fight against oxidative stress. Overall, the present study demonstrated that caffeine may impact bivalves, even at environmentally relevant concentrations, inducing oxidative stress in organisms. The present study is an important contribution to address knowledge gaps regarding the impacts of long-term exposures to pharmaceuticals since most of the studies assessed the effects after acute exposures, most of them up to 96 h. KW - Bivalves KW - Oxidative stress KW - Pharmaceuticals KW - Long-term exposure KW - Environmentally relevant concentrations PY - 2016 DO - https://doi.org/10.1016/j.chemosphere.2016.06.068 VL - 2016 IS - 160 SP - 95 EP - 103 PB - Elsevier Ltd. AN - OPUS4-38508 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Pires, A. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Schneider, Rudolf A1 - Esteves, V. I. A1 - Wrona, F. J. A1 - Soares, A. M. V. M. A1 - Figueira, E. A1 - Freitas, R. T1 - Hediste diversicolor as bioindicator of pharmaceutical pollution: Results from single and combined exposure to carbamazepine and caffeine N2 - Several environmental stressors have been identified as key and/or emerging drivers of habitat change that could significantly influence marine near-shore ecosystems. These include increasing discharges of pharmaceutical contaminants into the aquatic coastal systems. Pharmaceutical drugs are often detected in aquatic environments but still information on their toxicity impacts on inhabiting species is scarce, especially when acting in combination. Furthermore, almost no information is available on the impacts of pharmaceuticals in polychaetes, often the most abundant taxon in benthic communities and commonly used as indicator species of environmental conditions. Therefore, the present study aimed to evaluate the biochemical alterations induced in the polychaete Hediste diversicolor, from a low contaminated area at the Ria de Aveiro lagoon (Portugal), by the antiepileptic drug carbamazepine (0.0 - control, 0.3, 3.0, 6.0 and 9.0 μg/L) and the stimulant caffeine (0.0 - control, 0.5, 3.0, and 18.0 μg/L), acting alone and in combination (0.3 CBZ + 0.5 CAF and 6.0 CBZ + 3.0 CAF). Glutathione Stransferases (GSTs), superoxide dismutase (SOD) and catalase (CAT) activities was determined in Hediste diversicolor from each condition. Lipid peroxidation (LPO), glutathione reduced and oxidized (GSH and GSSG), glycogen and electron transport system (ETS) were also measured. The results obtained clearly revealed that both drugs induced oxidative stress in H. diversicolor, shown by the increase on LPO levels and decrease on total glutathione and GSH/GSSG ratio with the increase of exposure concentrations. Furthermore, the present findings demonstrated that polychaetes biotransformation capacity as well as antioxidant defense mechanisms were not sufficiently efficient to fight against the excess of reactive oxygen species (ROS) leading to LPO when organisms were exposed to both drugs. Our results also demonstrated that polychaetes tended to decrease the activity of ETSwhen exposed to drugs, avoiding energy expenditurewhich may prevent them fromgreater damages. The present study further revealed that the impacts induced by the combination of both drugswere similar to those obtained at the highest drugs concentrations acting alone. KW - Invertebrates KW - Pharmaceuticals KW - Oxidative stress biomarkers KW - Energy reserves PY - 2016 DO - https://doi.org/10.1016/j.cbpc.2016.06.003 VL - 2016 IS - 188 SP - 30 EP - 38 PB - Elsevier Inc. AN - OPUS4-38509 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schneider, Rudolf A1 - Teixiera, M. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Esteves, V. I. A1 - Wrona, F. J. A1 - Soares, A. M. V. M. A1 - Figueira, E. A1 - Freitas, R. T1 - Toxic effects of the antihistamine cetirizine in mussel Mytilus galloprovincialis N2 - Recent studies have become increasingly focused on the assessment of pharmaceuticals occurrence in aquatic ecosystems, however the potential toxicity to non-target organisms is still largely unknown. The antihistamine cetirizine is a commonly used pharmaceutical, already detected in surface waters of marine aquatic systems worldwide. In the present study Mytilus galloprovincialis mussels were exposed to a range of cetirizine concentrations (0.3, 3.0, 6.0 and 12.0 mu/L), resembling moderate to highly contaminated areas, over 28 days. The responses of different biochemical markers were evaluated in mussels whole soft tissue, and included energy-related parameters (glycogen content, GLY; protein content, PROT; electron transport system activity, ETS), and oxidative stress markers (superoxide dismutase activity, SOD; catalase activity, CAT; glutathione S-transferases activity, GSTs; lipid peroxidation levels, LPO; reduced (GSH) and oxidized (GSSG) glutathione content). The results obtained demonstrated that with the increase of exposure concentrations mussels tended to increase their energy reserves and maintain their metabolic potential, which was significantly higher only at the highest concentration. Our findings clearly revealed that cetirizine inhibited the activity of GSTs and although induced the activity of antioxidant enzymes (SOD and CAT) mussels were not able to prevent cellular damages observed through the increase of LPO associated to the increase of exposure concentrations. Thus, this study confirmed that cetirizine induces toxic effects in Mytilus galloprovincialis, which, considering their trophic relevance, wide use as bioindicator and wide spatial distribution of this species, can result in ecological and economic negative impacts at a large scale. KW - Bivalves KW - Biomarkers KW - Oxidative Stress PY - 2017 DO - https://doi.org/10.1016/j.watres.2017.02.032 SN - 0043-1354 VL - 114 SP - 316 EP - 326 AN - OPUS4-43302 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -