Sandwich Enzyme-Linked Immunosorbent Assay for the Detection of Bovine Blood in Animal Feed

Ofori, Jack Appiah; Hsieh, Yun-Hwa Peggy

doi:10.1021/jf070034r

Cited by 18 publications

(17 citation statements)

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“…Our group has previously reported a competitive enzyme‐linked immunosorbent assay (cELISA) for the quantitative detection of bovine blood in heat‐processed meat and feed (Rao and Hsieh 2008). This cELISA is based on monoclonal antibody (MAb), Bb1H9, which recognizes a 12 kDa thermal‐stable protein in ruminant (bovine, ovine, and goat) blood, and overcomes the shortcomings, such as lack of species and blood specificity and ineffectiveness for analyzing heat‐treated samples as the case is for existing methods as discussed previously (Ofori and Hsieh 2007; Rao and Hsieh 2008). Because of MAb Bb1H9's strong affinity for its antigen and the excellent selectivity for ruminant blood (both raw and heat‐treated), it has become important for further studies on this antibody and its antigenic protein in order to better define the specific use of MAb Bb1H9.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a 12 kDa Thermal‐Stable Antigenic Protein in Bovine Blood

Ofori

Hsieh

2011

Journal of Food Science

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MAb Bb1H9 represents the first antibody with the capacity to recognize bovine hemoglobin both in the absence and presence of the heme group, regardless of the heat treatment. MAb Bb1H9 can therefore be utilized in immunoassays by manufacturers and regulators to detect any ingredients containing hemoglobin or globin (hemoglobin without the heme group) in both raw and processed food and feed materials for product quality control and labeling law enforcement.

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Section: Introductionmentioning

confidence: 99%

Characterization of a 12 kDa Thermal‐Stable Antigenic Protein in Bovine Blood

Ofori

Hsieh

2011

Journal of Food Science

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“…Cooked and autoclaved bovine blood, plasma, serum, and RBCs were prepared as previously described (Ofori and Hsieh ). Raw (nonheated) samples were used as is.…”

Section: Methodsmentioning

confidence: 99%

“…We have previously reported on a sandwich enzyme‐linked immunosorbent assay (sELISA) for the detection of bovine blood in animal feed that can be used for the surveillance of bovine spongiform encephalopathy (BSE), commonly known as mad cow disease (Ofori and Hsieh ). This assay utilizes 2 monoclonal antibodies (mAbs), Bb6G12 and Bb3D6, that recognize a 60‐kDa thermally stable antigenic protein in bovine blood (Hsieh and others ).…”

Section: Introductionmentioning

confidence: 99%

“…This assay utilizes 2 monoclonal antibodies (mAbs), Bb6G12 and Bb3D6, that recognize a 60‐kDa thermally stable antigenic protein in bovine blood (Hsieh and others ). This antigenic protein is likely to be present in the plasma fraction of blood because of its strong positive reaction with spray‐dried bovine plasma (Hsieh and others ; Ofori and Hsieh ). It is scientifically important to establish the identity of this 60‐kDa thermal stable protein, as it may provide a basis for differentiating bovine blood from that of other species.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of a 60‐kDa Thermally Stable Antigenic Protein as a Marker for the Immunodetection of Bovine Plasma‐Derived Food Ingredients

Ofori

Hsieh

2015

Journal of Food Science

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A sandwich enzyme-linked immunosorbent assay (sELISA) based on 2 monoclonal antibodies (Bb3D6 and Bb6G12) that recognize a 60-kDa antigenic protein in bovine blood was previously developed for detecting bovine blood in animal feed for the prevention of mad cow disease. This study sought to establish the identity of this 60-kDa antigenic protein and consequently determine the suitability of the sELISA for detecting bovine plasma-derived food ingredients (BPFIs), which are widely used in dietary products without explicit labeling. Results from western blot confirmed the 60-kDa protein to be present in the plasma fraction of bovine blood. Further proteomic analyses involving 2-dimensional gel electrophoresis (2-D GE) and amino acid sequencing revealed the 60-kDa protein to be bovine serum albumin (BSA). The sELISA proved capable of detecting BPFIs in all the commercial dietary supplements tested, including those that were formulated with hydrolyzed BPFIs. The assay could also detect 0.01% and 0.5% of different BPFIs in spiked raw and cooked ground beef, respectively. This assay based on the detection of BSA therefore has the potential to become a valuable analytical tool to protect consumers who avoid consuming BPFIs for religious, health, or ethical reasons.

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“…A wide variety of analytical methods for DEPAMEDE Media Peternakan the determination of pig components in foods or diet have been described. These include, methods based on electrophoresis (Kim & Shelef, 1986), isoelectric focusing (Jaussen et al, 1990), chromatography (Saeed et al, 1989), enzyme-linked immunosorbent assay (ELISA) (Hsieh et al, 1996;Chen et al, 1998;Macedo-Silva Macedo-Silva et al, 2000;Ofori & Hsieh, 2007;Asensio et al, 2008), electronic nose and ), electronic nose and gas chromatography mass spectrometer with headspace analyzer (GCMS-HS) (Nurjuliana et al, 2011), also fouri-fourier transform infrared (FTIR) spectroscopy (Rohman et al, 2011). In addition to those methods, methods based on In addition to those methods, methods based on DNA technology such as DNA hybridization (Ebbehøj & Thomsen, 1991;Ballin et al, 2009), polymerase chain reaction (PCR) (Matsunaga et al, 1995;Rensen et al, 2006;Rojas et al, 2009;Ballin et al, 2009;Rojas et al, 2010) have been used for more than two decades.…”

Section: Introductionmentioning

confidence: 99%

Development of a Rapid Immunodiagnostic Test for Pork Components in Raw Beef and Chicken Meats: a Preliminary Study

Depamede

2011

med pet

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ABSTRAKMetode uji cepat imunodiagnostik untuk memastikan adanya cemaran komponen daging babi pada daging sapi dan daging ayam mentah telah berhasil dikembangkan pada penelitian ini. Prinsip pengembangan imunodiagnostik ini adalah dengan mengkonjugasikan koloid emas sebagai penanda dengan antibodi poliklonal IgG babi. Konjugat ini diletakkan pada bantalan konjugat, salah satu bagian dari sistem "strip tes" imunokromatografi. Strip tes yang dihasilkan kemudian digunakan untuk uji ada tidaknya cemaran komponen daging babi. Ekstrak campuran daging mentah yang digiling halus dalam bufer garam fosfat, berupa campuran daging babi/daging sapi atau daging babi/daging ayam dalam perbandingan 1/0; 1/100; 1/1000; 1/5.000; 1/10.000 (berat/berat) digunakan pada studi pendahuluan dalam skala laboratorium. Sebagai kontrol digunakan ekstrak daging sapi dan daging ayam mentah, tanpa campuran daging babi sama sekali. Hasil penelitian menunjukkan bahwa dalam waktu 10 menit cemaran komponen daging babi dapat dideteksi secara kasat mata pada campuran 1 bagian daging babi per 5.000 bagian daging sapi atau daging ayam mentah, bahkan pada campuran 1/10.000 masih dapat terdeteksi. Tes immunodiagnostik ini dapat diterapkan untuk mendeteksi dengan mudah komponen daging babi pada produk olahan daging sapi atau daging ayam mentah. Perlu dilakukan penelitian lebih lanjut untuk pengembangan skala komersial. Kata kunci: pencemaran daging, imunodiagnostik, daging babi, uji cepat ABSTRACTA rapid immunodiagnostic test that provides visual evidence of the presence of pork components in raw beef and chicken meats was developed. Colloidal gold was prepared and conjugated with anti-Swine IgG polyclonal antibody. Immunochromatographic test strips were produced, and then were used to test laboratory adulterated raw meat samples. The samples consisted of pork-inbeef, or pork-in-chicken at 1/0; 1/100; 1/1,000; 1/5,000; 1/10,000 (w/w) adulteration levels that were extracted in phosphate-buffered saline. Raw beef and chicken meats without pork were included as controls. Analysis was completed in 10 min. Detection limit was 1/5,000 (w/w), although 1/10,000 was also observed. This immunodiagnostic tests can be conveniently applied to detect low levels of pork components in raw beef and chicken meat products. For the commercial purposes, further studies need to be carried out.

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Sandwich Enzyme-Linked Immunosorbent Assay for the Detection of Bovine Blood in Animal Feed

Abstract: Bovine; blood; monoclonal antibody; sandwich ELISA.

Cited by 18 publications

References 16 publications

Characterization of a 12 kDa Thermal‐Stable Antigenic Protein in Bovine Blood

Characterization of a 12 kDa Thermal‐Stable Antigenic Protein in Bovine Blood

Characterization of a 60‐kDa Thermally Stable Antigenic Protein as a Marker for the Immunodetection of Bovine Plasma‐Derived Food Ingredients

Development of a Rapid Immunodiagnostic Test for Pork Components in Raw Beef and Chicken Meats: a Preliminary Study

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