Evaluation of a Commercial ELISA for Detection of Ruminant Processed Animal Proteins in Non-Ruminant Processed Animal Proteins
Bremer, M.G.E.G. ; Margry, R.J.C.F. ; Vaessen, J.C.H. ; Doremalen, A.M.H. van; Palen, J.G.P. van der; Kaathoven, R.G.C. van; Kemmers-Voncken, A.E.M. ; Raamsdonk, L.W.D. van - \ 2013
Journal of AOAC International 96 (2013)3. - ISSN 1060-3271 - p. 552 - 559.
plant sterilization conditions - shrimp litopenaeus-vannamei - linked-immunosorbent-assay - bone meals - classical microscopy - rendered meat - by-products - feed - pcr - identification
Due to a growing aquaculture industry, demand for high-quality proteins for aquatic feeds is increasing. Non-ruminant processed animal proteins (PAPs) have shown great potential for this purpose. Safe reintroduction of non-ruminant PAPs in aqua feed requires methods that can discriminate ruminant and non-ruminant PAPs at contamination levels at or below 2%. Because the official European Union method lacks species specificity, the performance of MELISA-TEK™ Ruminant, a commercial immunoassay, combined with the MELISA-TEK High Sensitivity Sample Extraction kit was evaluated. Various non-ruminant PAPs spiked with ruminant PAPs (processed at 133, 137, 141, and 145°C) were analyzed. Results showed an overall specificity of 99%, indicating no cross-reaction with non-ruminant PAPs. The sensitivity of the assay strongly depended on both processing temperature and proportion of muscle fibers of the ruminant PAPs. Overall sensitivity of samples with 1 and 2% ruminant PAPs was 92 and 100%, respectively. For ruminant PAPs processed at 133 and 137°C, the sensitivity was 100% for both 1 and 2% ruminant spikes. Overall accuracies were 96 and 99% for 1 and 2% ruminant spikes, respectively. In conclusion, the MELISA-TEK Ruminant assay showed satisfactory results, which makes it a suitable candidate method to enable safe reintroduction of non-ruminant PAPs in aqua feed.
New developments in classical microscopy; what can be expected for the official control?
Raamsdonk, L.W.D. van; Pinotti, L. ; Veys, P. ; Bremer, M.G.E.G. ; Hekman, W.E. ; Kemmers-Voncken, A.E.M. ; Campagnoli, A. ; Paltanin, C. ; Crespo, C. ; Vliege, J.J.M. ; Pinckaers, V.G.Z. ; Jorgensen, J.S. - \ 2011
Biotechnology, Agronomy, Society and Enviroment 15 (2011)s1. - ISSN 1370-6233 - p. 15 - 24.
bovine spongiform encephalopathy - pcr detection - by-products - feeds
The official control of animal proteins in feed is focused on the prevention of Bovine Spongiform Encephalopathy (mad cow disease). The current legislation of the European Union is planned to avoid the feeding of animal by-products to the same species as its origin (ban of cannibalism, or species-to-species ban). With respect to the official control, the circumscription of the term species in legislation should be defined, and species-specific markers should be available. Markers will include primer sets, antibodies, near-infrared profiles or visual characteristics. The method of classical light microscopy is currently the only accepted method in the framework of the official detection of animal proteins. Besides the necessary development of complementary methods, either as stand alone methods or in combination, the visual characteristics used for a microscopic examination of meat and bone meal particles should be fully explored. Multivariate analysis of a range of characteristics of lacunae in bone fragments revealed that discrimination is possible between mammalian and avian bone fragments. Translation to features for every day practical use should be carried out very carefully, and only comprehensively collected information on a range of features will give a first indication of the source. Characteristics of hairs and feather filaments can be used to identify the origin of animal particles. An in situ identification method has been developed for antibody conjugation with troponin I in muscle fibers on a microscopic slide. A proof of principle is presented. Interlaboratory transferability and validation have still to be achieved. The development and testing of light microscopy markers in the framework of the SAFEED-PAP project revealed that a fine tuning of existing microscopic characteristics appears to be possible. Keywords. Microscopic method, feed, marker, hair, feather, bone, meat and bone meal.
Immunochromatographic Lateral-flow test strip for the rapid detection of added bovine rennet whey in milk and milk powder
Martin-Hernandez, C. ; Munoz, M. ; Daury, C. ; Weymuth, H. ; Kemmers-Voncken, A. ; Corbation, V. ; Toribo, T. ; Bremer, M.G.E.G. - \ 2009
International Dairy Journal 19 (2009)4. - ISSN 0958-6946 - p. 205 - 208.
reversed-phase hplc - buttermilk powder - skim milk - uht milk - capillary-electrophoresis - solids - proteolysis
An immunochromatographic lateral-flow test dipstick test was developed for the fast detection of bovine rennet whey in liquid milk and milk powder. The test is based on the binding of casein glycomacropeptide (cGMP) by two specific anti-bovine ¿-casein monoclonal antibodies and has a visual detection limit of around 15 ng mL¿1 for cGMP and 1% (v/v) for rennet whey in milk using standards and spiked samples. The dipstick performance was evaluated in a collaborative trial using skim milk powder and raw, pasteurized and UHT milk. The suitability of the dipstick was demonstrated in comparison with gel permeation-high performance liquid chromotography and a colorimetric method, by analysing 60 raw milk samples collected from farms in Brazil. The dipstick results correlated well with the HPLC results and were more reliable than those obtained with the colorimetric method. The dipstick test correctly identified all raw milk samples with a rennet whey content above 4%.
Enzyme-linked immunosorbent assay for the detection of bovine rennet whey powder in milk powder and buttermilk powder
Bremer, M.G.E.G. ; Kemmers-Voncken, A. ; Boers, E.A.M. ; Frankhuizen, R. ; Haasnoot, W. - \ 2008
International Dairy Journal 18 (2008)3. - ISSN 0958-6946 - p. 294 - 302.
performance liquid-chromatography - capillary-zone-electrophoresis - kappa-casein - biosensor immunoassays - dairy-products - caseinomacropeptide - solids - glycomacropeptide - macropeptide - temperature
An inhibition enzyme-linked immunosorbent assay (ELISA) for the detection of bovine rennet whey (BRW) solids in skim milk powders (SMP) and buttermilk powders is presented. The BRW content was determined in a neutralised trichloroacetic acid sample extract by binding of the dissolved caseinomacropeptide to an enzyme-labelled anti-bovine-¿-casein monoclonal antibody. Calibration curves were constructed by analysing SMP standards with different known concentrations of BRW (0¿5.8% (w/w)).The assay has a limit of detection of 0.1% (w/w) BRW powder in SMP and has high repeatability and reliability. The ELISA reached the sensitivity required for screening to conform with European Union (EU) legislation. It is easy to use, has a short assay time and is of low cost. The successful applicability of this new screening assay was demonstrated in comparison with chromatographic methods imposed by the EU, where 60 industrial samples taken by the Dutch General Inspection Service were analysed.
Detection of hidden hazelnut protein in food by IgY-based indirect competitive enzyme-immunoassay
Baumgartner, S. ; Bremer, M.G.E.G. ; Kemmers - Voncken, A.E.M. ; Smits, N.G.E. ; Haasnoot, W. ; Banks, J. ; Reece, P. ; Danks, C. ; Tomkies, V. ; Immer, U. ; Schmitt, K. ; Krska, R. - \ 2004
Analytica Chimica Acta 520 (2004)1/2. - ISSN 0003-2670 - p. 223 - 228.
affinity-chromatography - anaphylaxis - antibodies - selection
The development of an indirect competitive enzyme-immunoassay for the detection of hidden hazelnut protein in complex food matrices is described. A sensitive and selective polyclonal antibody was raised by immunisation of laying hens with protein extracts from roasted hazelnuts. In contrast to traditional antibody generation in mammals, the antibody was not isolated from the blood of immunised mammals but from the egg yolk of immunised chickens. A standard calibration curve was optimised using immunoaffinity purified antibody extract and a coating antigen concentration of 10 ¿g ml¿1. One percent skim milk powder was chosen for blocking. The assay has a minimum detection limit of 10 ¿g l¿1, with an IC50 of 618 ¿g l¿1 when a 50 mM phosphate buffer at pH 7.5 and 10 mM sodium chloride is used as assay buffer. The cross reactivity testing shows a high specificity for hazelnut proteins and various foods and food additives were found to be non reactive except beans, sunflower seed or poppy seed.
Fast biosensor immunoassays for the detection of cows' milk in the milk of ewes and goats
Haasnoot, W. ; Smits, N.G.E. ; Voncken, A.E.M. ; Bremer, M.G.E.G. - \ 2004
Journal of Dairy Research 71 (2004)3. - ISSN 0022-0299 - p. 322 - 329.
linked-immunosorbent-assay - bovine beta-casein - monoclonal-antibodies - polyclonal antibodies - liquid-chromatography - k-casein - cheese - elisa - proteins - ovine
Two monoclonal antibodies (MAb) raised against bovine K-casein were developed and applied in an automated optical biosensor (Biacore 3000) to create easy and fast direct and inhibition biosensor immunoassays (BIA) for the detection of cows' milk in the milk of ewes and goats. With both assay formats, low limits of detection (350 tests); and the wide measurement range (0·1 to 10% cows' milk). Despite these advantages, the inhibition BIA (using K-casein immobilized on the chip) was preferred because of the possible application of non-purified Mab, the higher responses, the higher sensitivity at relevant low percentages of cows' milk and its robustness (>800 cycles per chip)
|Medroxyprogesterone acetate: development and validation of a screening method for porcine fat
Made, E.A.J. van der; Haasnoot, W. ; Voncken, A.E.M. ; Keukens, H.J. - \ 2004
In: Proceedings of the EuroResidue V conference, Noordwijkerhout 10-12 May 2004 Utrecht : University of Utrecht - p. 644 - 648.
|Rapid tests for allergen detection
Bremer, M. ; Kemmers-Voncken, A. ; Smits, N.G.E. ; Haasnoot, W. ; Baumgartner, S. ; Krska, R. ; Banks, J. ; Reece, P. ; Danks, C. ; Tomkies, V. ; Schmitt, K. ; Immer, U. ; Kiening, M. ; Corsini, E. ; Wilson, P. ; Scarniet, I. - \ 2003
In: 3rd symposium on 'Product safety, quality and competitiveness in the food sector', 6, 7 & 8 November 2003 [S.l.] : S.n. - p. 482 - 492.
Immunofiltration as sample cleanup for the immunochemical detection of beta -agonists in urine
Haasnoot, W. ; Kemmers-Voncken, A. ; Samson, D. - \ 2002
The Analyst 127 (2002)1. - ISSN 0003-2654 - p. 87 - 92.
Despite the ban of the European Union on use of drugs to improve animal growth, occasionally -agonist drugs are still found in samples from cattle. Over time, the specified limits for the detection of these illegal drugs have been lowered. To improve the immunochemical screening of urine samples to detect lower levels of several -agonists, immunofiltration (IF) was applied for sample cleanup in combination with a -agonist-ELISA. In the applied IF format, free (non-immobilised) anti-salbutamol polyclonal antibodies were mixed with the urine sample in an ultra-filtration device (cut off 30 kDa) and the sample was removed by centrifugation. The antibody bound -agonists were freed from the antibodies by the addition of a mixture of methanol and 0.1 M acetic acid (11; v/v) and centrifugation. The filtrate, containing the free -agonists, was evaporated to dryness and the residue dissolved in buffer, an aliquot of which was analysed with the -agonist ELISA. Compared with the direct -agonist ELISA, this IF cleanup procedure resulted in a 30-times lower limit of detection (LOD) of 0.14 ng ml¿1 (salbutamol equivalents). The anti-salbutamol antibodies recognised several -agonists and the combination of IF with the -agonist ELISA was found suitable for the detection of at least ten -agonists in urine with comparable LODs.
|Immunofiltration as alternative for immunoaffinity chromatography
Haasnoot, W. ; Kemmers-Voncken, A. ; Rhijn, H. van; Schilt, R. - \ 2000
In: Proceedings of the EuroResidue IV Conference, Veldhoven, The Netherlands
|Application of generic monoclonal antibodies against sulfonamides in optical biosensors
Haasnoot, W. ; Kohen, F. ; Pre, J. du; Cazemier, G. ; Kemmers-Voncken, A. ; Bienenmann-Ploum, M. ; Verheijen, R. - \ 2000
In: Proceedings of the EuroResidue IV Conference, Veldhoven, The Netherlands
Sulphonamide Antibodies: From Specific Polyclonals to Generic Monoclonals
Haasnoot, W. ; Cazemier, G. ; Pre, J. du; Kemmers-Voncken, A. ; Bienenmann-Ploum, M. ; Verheijen, R. - \ 2000
Food and Agricultural Immunology 12 (2000). - ISSN 0954-0105 - p. 15 - 30.
Monoclonal antibodies against a sulfathiazole derivative for the immunochemical detection of sulfonamides
Haasnoot, W. ; Pre, J. Du; Cazemier, G. ; Kemmers-Voncken, A. ; Verheijen, R. ; Jansen, B.J.M. - \ 2000
Food and Agricultural Immunology 12 (2000). - ISSN 0954-0105 - p. 127 - 138.
To prepare monoclonal antibodies (mAbs) against the generic part of sulfonamides, a sulfathiazole derivative was chemically linked to carrier proteins in such a way that the aromatic amino group, common to all sulfonamides, was distal to the proteins. Four mice were immunized with the sulfathiazole-protein derivatives. The spleen cells of one of the mice were fused with myeloma cells to produce hybridomas of which the supernatants were screened in an indirect ELISA (iELISA) for the presence of sulfathiazole antibodies. After cloning, positive supernatants were tested in a competitive iELISA (ciELISA) for inhibition with 18 sulfonamides. This resulted in four different mAbs (all IgG1 kappa light chain) which recognized several sulfonamides. By use of the best monoclonal (27G3) and an optimized ciELISA protocol, eight structurally different sulfonamides showed 50% inhibition at concentrations less than 100 ngml-1 or 5 ng/well. However, other relevant sulfonamides (such as sulfadimidine, sulfatroxazole and sulfachloropyrazine) were detected at a high level only with this mAb. This means that the ciELISA (with the best Mab) showed a broad specificity for sulfonamides but the sensitivity towards the different sulfonamides varied too much to call it a generic sulfonamide ELISA.
|Hygiene Motivation in India and the Netherlands
Curtis, V. ; Cave, B. ; Voncken, N. ; Singh, S. ; Niehof, A. ; Butijn, C. - \ 1999
London : London School of Hygiene and Tropical Medicine - 71 p.
|Hygiene in the Netherlands; An inquiry into ideas and habits of people regarding hygiene (Abridged Report)
Niehof, A. ; Voncken, N. - \ 1999
Wageningen : Wageningen Agricultural University, Household and Consumer Studies - 37
hygiëne - moeders - moederlijk gedrag - nederland - persoonlijke hygiëne - hygiene - mothers - maternal behaviour - netherlands - personal hygiene
|Immunochemical approaches to the analysis of ß-agonistic drugs
Haasnoot, W. ; Cazemier, G. ; Stouten, P. ; Kemmers-Voncken, A. - \ 1996
In: ACS Symposium Series 621. Immunoassays for residue analysis, Anaheim, Calif., 2-7 april 1995, Eds R.C. Beier and L.H. Stanker, ed. 1996 - p. 60 - 73.
|Als je toch thuis bent.
Casimir, G.J. - \ 1996
In: Flexibilisering in arbeid en loopbaan. Studiedag Flexibilisering in arbeid en loopbaan. KLV Wageningen / Voncken, P., - p. 55 - 60.
Ontwikkeling van een veldtest voor de bepaling van ß-agonisten in urine
Haasnoot, W. ; Streppel, L. ; Cazemier, G. ; Salden, M. ; Stouten, P. ; Kemmers Voncken, A. ; Wichen, P. van - \ 1995
Wageningen : DLO-Rijks-Kwaliteitsinstituut voor Land- en Tuinbouwprodukten - 22
bèta-adrenerge agonisten - vleesproductie - dierlijke producten - urine-analyse - urine - beta-adrenergic agonists - meat production - animal products - urine analysis - ß-agonists - on-site test - enzyme immunoassay
Mouse models in leukemia
Voncken, J.W. - \ 1995
Agricultural University. Promotor(en): R.W. Goldbach; J.H.C. Groffen. - S.l. : Voncken - ISBN 9789054854166 - 148
gammaretrovirus - virologie - moleculaire biologie - leukemie - muridae - muizen - modellen - onderzoek - gammaretrovirus - virology - molecular biology - leukaemia - muridae - mice - models - research
Human Philadelphia-positive leukemia results from a balanced chromosomal translocation, which fuses the BCR gene on chromosome 22 to the ABL proto-oncogene on chromosome 9. The understanding of Ph-positive leukemogenesis has advanced enormously over the last few decades. Although in vitro assay systems currently used, are not always relevant to human tumor biology, much can and has been learned from studies, employing cell cultures and overexpression of BCR/ABL oncogenes.
Another restriction in leukemia research is the availability of primary human tumor material for study. Moreover, such tissues often represent terminally advanced stages of tumorigenesis. Therefore, the importance of in vivo models to study Philadelphia-positive leukemia is manifold. A well defined transgenic mouse model allows for tumorigenesis to be studied from its earliest stages onward and factors and mechanisms that eventually contribute to malignant progression of the leukemic cells can be uncovered. Besides an 'unlimited' provision of tumor material for analysis, more importantly, the availability of a transgenic mouse model provides a means by which cancer treatment regimes can be tested. In addition, identification of cellular components and/or pathways that contribute to the onset or progression of leukemia may eventually lead to the discovery and development of new drugs.
In 1990, Heisterkamp and co-workers reported on a transgenic mouse model for Philadelphia-positive acute lymphoblastic leukemia (ALL). Since most of the transgenic animals of an earlier study had succumbed to leukemia, part of the aim of this thesis was to generate BCR/ABL P190 transgenic founder animals de novo and to derive a transgenic animal line(s) which was to be used for future studies. In order to better understand the animal model, leukemogenesis was studied in great detail in transgenic founder animals and their progeny. In the second chapter a cytogenetic study of the mouse model for acute lymphoblastic leukemia is presented. Karyotypic analysis of leukemic bone marrow of a significant number of mice shows, that leukemic cells undergo a clonal development and karyotype evolution toward a more aggressive tumor: a high frequency of aneuploidy is found in advanced leukemia, as occurs in human leukemia, with a preference for gain of chromosomes 10, 12, 14 and 17. These findings are corroborated by experiments that reveal a gain of malignancy of the cancer upon serial transplantation of leukemic bone marrow to irradiated recipient mice and by molecular analysis of lymphomas using immunoglobulin rearrangement as an indicator for tumor clonality. The results suggest that BCR/ABL has a destabilizing effect on the regulation of the proces of mitosis.
In the third chapter, a correlation is described between the transcriptional status of the BCR/ABL P190 transgene and the development of leukemia: methylation of particular sequences in BCR exon-1 in the transgene is closely coupled to transgene inactivation, providing additional evidence for a direct role of BCR/ABL in leukemogenesis. A biological dissection of the oncogenic specificity of BCR/ABL is presented in the fourth chapter. Using sensitive molecular biological techniques, it is shown that, although expression of the BCR/ABL transgene is detectable in every tissue, from very early on in mouse development, no other neoplasias than of hematopoietic origin are found. The results strongly suggest that the oncogenicity of BCR/ABL is restricted to nucleated blood cells, which is very likely a reflection of cellular functions of the BCR and or ABL gene in signal transduction specific to hematopoietic lineages. The observations would also explain why the Ph-chromosome, which one would expect to arise by chance in many proliferating tissues, is found only in blood cancers.
An analysis of transgenic mouse models for chronic myelogenous leukemia, using BCR/ABL P210 transgenes is presented in the fifth chapter. The clinical disease spectrum includes differentiated and undifferentiated T and B cell leukemias. The myeloid compartment is implicated only sporadically and rather late in the disease process. In some instances, the observed myelo-proliferation is a sequel to deregulation of cytokine production at advanced stages of leukemia. The course of P210 induced leukemia was acute rather than chronic, be it with an on average longer latency period than typical for ALL in BCR/ABL P190 mice. From these studies is was concluded that in the mouse, BCR/ABL P210 evokes a clinically different disease than BCR/ABL P190. Although no evidence for a chronic myeloproliferative disorder in the peripheral blood was found, an imbalance in myelopoiesis in the bone marrow suggests an effect of BCR/ABL P210 on primitive myeloid progenitors.
The sixth chapter summarizes an analysis of interferon-α(IFN-α) treatment of the BCR/ABL P190 transgenic mice. (IFN-α) is currently one of the most effective drugs in the treatment of CML. Recently, (IFN-α) was tried in the treatment of ALL. No effect of (IFN-α) on animal survival or disease pattern were noted when administered to the BCR/ABL P190 mice. The conclusion was reached that, at least in a transgenic setting, (IFN-α) does not interfere with BCR/ABL P190 mediated leukemia.
In order to study the normal cellular function of the BCR gene and to eventually assess its role in leukemogenesis, studies focussing on the mouse bcr gene function are presented in chapters 7 and 8. The seventh chapter describes the ablation of a functional mouse bcr gene by means of recently developed gene targeting techniques. One of two mouse bcr alleles was inactivated in a mouse embryonic stem cell line through gene interruption by insertional replacement vectors. Ibis genetically altered cell fine was then injected into developing mouse embryos. Through germline transmission of the mutated allele and subsequent breeding both bcr alleles were inactivated. Although bcr -null mutants are phenotypically normal, their neutrophils display impaired regulation of respiratory burst, which becomes apparent when these cells are activated in vivo: an overproduction of superoxide leads to significantly more oxidative tissue damage during experimental endotoxemia. The results connect Bcr in vivo with the regulation of superoxide production by the NADPH- oxidase system of leukocytes and suggest a link between the cell types affected by loss of Bcr function and the those involved in Ph -positive leukemia.
Additional information on biological processes that Bcr participates in, are described. in the eighths chapter. Notwithstanding its function in hematopoietic cells, the Bcr protein is normally found in high levels in brain. The expression pattern of bcr in rodent brain was examined by means of in situ hybridization and Northern analysis. Although not directly connected with leukemogenesis, a potentially interesting role for p160Bcr in the brain is discussed as its expression pattern appears to coincide with the functional organization of particularly highly specialized structures in the brain.
With the availability of well defined transgenic mouse models for BCR/ABL positive leukemia, an opportunity is created to study the nature of cellular interactions and processes that contribute to the onset and development of Ph -positive leukemia. Ultimately, such investigations are aimed at designing and testing effective therapeutic drugs to fight the disease.
|Detection and analysis of Autographa californica nuclear polyhedrosis virus mutants with defective interfering properties.
Kool, M. ; Voncken, J.W. ; Lier, F.L.J. van; Tramper, J. ; Vlak, J.M. - \ 1991
In: Abstract 11th Meeting Eur. Soc. Animal Cell Technology, Brighton, UK - p. 65 - 65.