Application of Immunoaffinity Column as Cleanup Tool for an Enzyme Linked Immunosorbent Assay of Phosphinothricin-N-acetyltransferase Detection in Genetically Modified Maize and Rape

2005 ◽  
Vol 53 (11) ◽  
pp. 4315-4321 ◽  
Author(s):  
Wentao Xu ◽  
Kunlun Huang ◽  
Heng Zhao ◽  
Yunbo Luo
Keyword(s):  
Immunosorbent Assay ◽  
Genetically Modified ◽  
Enzyme Linked Immunosorbent Assay ◽  
Genetically Modified Maize ◽  
Immunoaffinity Column

Development of an Enzyme-Linked Immunosorbent Assay and Immunoaffinity Column Chromatography for Saikosaponin d Using an Anti-Saikosaponin d Monoclonal Antibody

Planta Medica ◽  
2016 ◽  
Vol 82 (05) ◽  
pp. 432-439 ◽  
Author(s):  
Jiayang Sai ◽  
Yan Zhao ◽  
Wenchao Shan ◽  
Baoping Qu ◽  
Yue Zhang ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Column Chromatography ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunoaffinity Column

Enzyme linked immunosorbent assay for PAT protein detection in genetically modified rape

2006 ◽  
Vol 3 (3) ◽  
pp. 177-181 ◽  
Author(s):  
Xu Wen-Tao ◽  
Huang Kun-Lun ◽  
Deng Ai-Ke ◽  
Luo Yun-Bo
Keyword(s):  
Immunosorbent Assay ◽  
Polyclonal Antibody ◽  
Genetically Modified ◽  
Protein A ◽  
Protein Detection ◽  
Cross Reactivity ◽  
Enzymic Activity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sepharose 4B ◽  
Pat Protein

AbstractWe have developed and applied an immunoassay method to detect genetically modified (GM) rape containing phosphinothricin acetyltransferase (PAT). The purified PAT was identified by Western blotting and enzymic activity analysis. The polyclonal antibody against purified PAT protein was obtained and purified by both a saturated ammonium sulphate method and protein A-Sepharose 4B. The sensitivity and cross-reactivity of the polyclonal antibody has been demonstrated in an enzyme-linked immunosorbent assay (ELISA). The result of the ELISA for antiserum sensitivity was about 2×10−5mg/ml and the cross-reactivity determined experimentally showed a high degree of specificity for the antiserum used, because values were all less than 0.1%. Detection of transgenic plants was evaluated using two transgenic rape lines (MS1/RF1 and MS8/RF3) which could be easily distinguished by ELISA.

scholarly journals Alergenisitas Sistem Glikasi Isolat Protein Kedelai-Fruktooligosakarida (Allergenicity Properties of Soy Protein Isolate-Fructooligosaccaride Glycation Systems)

2017 ◽  
Vol 36 (4) ◽  
pp. 450 ◽  
Author(s):  
Rahayu Suseno ◽  
Nurheni Sri Palupi ◽  
Endang Prangdimurti
Keyword(s):  
Immunosorbent Assay ◽  
Soy Protein ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Soy Protein Isolate ◽  
Immunological Response ◽  
Liquid System ◽  
Protein Isolate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Genetically Modified Food

Food allergy is an immunological response caused by allergens contained in food. Soybean is one of the eight kinds of food products that can cause allergies. Genetically modified food crops that are most widely produced worldwide is soybean (47 % worldwide). Genetically Modified Organisms (GMO) products is concerned may increase the allergenicity of the  product. The aims of the research were to study the allergenicity of GMO and non-GMO Soy Protein Isolates (SPI) and the glycation effect to allergenicity of SPI. GMO and non-GMO SPI were glycated with fructooligosaccharides (FOS) through the Maillard reaction in liquid systems. Allergenicity was determined qualitatively using immunoblotting and quantitatively using Enzyme-Linked Immunosorbent Assay (ELISA). The glycation degree of GMO and non-GMO SPI can increase up to 75.03 % and 73.50 % in the liquid system. There were 9 protein allergens in GMO soybean and 8 protein allergens in non-GMO soybean. The glycation reaction could reduce protein allergens in GMO and non-GMO SPI up to 91.69 % and 87.07 %.ABSTRAKAlergi pangan merupakan sebuah respon imunologis yang disebabkan oleh alergen yang terdapat pada pangan. Kacang kedelai merupakan satu dari delapan jenis bahan pangan yang sering menyebabkan alergi. Tanaman pangan hasil rekayasa genetika (GMO) yang banyak diproduksi di dunia adalah kacang kedelai yaitu sekitar 47 %. Produk GMO dikhawatirkan dapat meningkatkan alergenisitasnya. Penelitian ini bertujuan untuk mempelajari tinggat alergenisitas antara Isolat Protein Kedelai (IPK) GMO dan non-GMO serta pengaruh glikasi terhadap alergenisitas IPK. IPK GMO dan non-GMO diglikasi dengan fruktooligosakarida melalui reaksi Maillard dengan sistem cair. Alergenisitas diukur secara kualitatif menggunakan immunobloting dan secara kuantitatif menggunakan Enzyme-Linked Immunosorbent Assay (ELISA). Peningkatan derajat glikasi IPK GMO dan non-GMO pada sistem cair masing-masing memperlihatkan hasil 75,03 % dan 73,50 %. Terdapat 9 protein alergen pada kacang kedelai GMO dan 8 protein alergen pada kacang kedelai non-GMO. Reaksi glikasi dapat mengurangi alergen pada kacang kedelai GMO dan non-GMO hingga 91,69% dan 87,07%.

Screening of genetically modified organisms and specific detection of Bt176 maize in flours and starches by PCR-enzyme linked immunosorbent assay

2003 ◽  
Vol 217 (1) ◽  
pp. 83-89 ◽  
Author(s):  
Laetitia Petit ◽  
Fabienne Baraige ◽  
Anne-Marie Balois ◽  
Yves Bertheau ◽  
Patrick Fach
Keyword(s):  
Immunosorbent Assay ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Detection

scholarly journals Immunoaffinity Column as Cleanup Tool for a Direct Competitive Enzyme-Linked Immunosorbent Assay of Cyclopiazonic Acid in Corn, Peanuts, and Mixed Feed

1998 ◽  
Vol 81 (6) ◽  
pp. 1169-1176 ◽  
Author(s):  
Wanjun Yu ◽  
Joe W Dorner ◽  
Fun S Chu
Keyword(s):  
Immunosorbent Assay ◽  
Binding Capacity ◽  
Cyclopiazonic Acid ◽  
Chromatographic Behavior ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunoaffinity Column ◽  
Mixed Feed ◽  
Buffer Solution ◽  
Sample Extract ◽  
Sepharose 4B

Abstract An immunoaffinity column (IAC) for cyclopiazonic acid (CPA) was prepared by coupling a CPA-specific monoclonal antibody to CNBr-activated sepharose 4B. A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) was used to study the chromatographic behavior of a 0.2 ml_ gel column with a binding capacity of 4 μg CPA/column as well as to evaluate its efficacy as a cleanup tool for analysis of naturally occurring CPA. Sample extract either in buffer solution or in a solution containing up to 35% methanol could be loaded onto the column. After the column is washed with 5 ml_ deionized water and 5 ml_ 50% methanol, CPA could be quantitatively eluted with 2 ml_ 100% methanol. The column could be regenerated at least 10 times by washing with 10 mL equilibrating buffer and then storing in a cold room overnight before reuse. Recoveries of CPA added to corn, peanut, and mixed feed extracts in the range 10-200 ng/g were 88-105,86-100, and 90-110%, respectively. Detection limits were 2.0,4.4, and 4.7 ng/g for corn, mixed feed, and peanuts, respectively. Twenty-two peanut samples naturally contaminated with CPA were subjected to both IAC and solvent partition cleanup followed by dc-ELISA. Although a good correlation between data obtained from lAC-dc-ELISA and from SP-dc-ELISA (r = 0.75, p < 0.0001) was obtained, the slope of the linear regression was low (0.67), indicating loss during solvent partition cleanup. The overall data showed that the combination of IAC and dc- ELISA is an effective method for CPA analysis.

scholarly journals Quantitation of Aflatoxin B1-N7-Guanine Adduct in Urine by Enzyme-Linked Immunosorbent Assay Coupled with Immunoaffinity Chromatography

1997 ◽  
Vol 80 (5) ◽  
pp. 1013-1022 ◽  
Author(s):  
Tfflrumurthy Vidyasagar ◽  
Vadapalli Vyjayanthi ◽  
Nayak Sujatiia ◽  
Beedu Sashidhar Rao ◽  
Ramesh Venkataramana Bhat
Keyword(s):  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Polyclonal Antibodies ◽  
Immunoaffinity Chromatography ◽  
Competitive Elisa ◽  
Aflatoxin M1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunoaffinity Column ◽  
Indirect Competitive Elisa

Abstract IndiaA specific and sensitive method to quantitate aflatoxin B1-N7-guanine adduct in urine samples by immunoaffinity chromatography coupled with indirect competitive enzyme-linked immunosorbent assay (ELISA) is reported. A novel in vitro method to synthesize an antigen (bovine serum albumin-guanine-aflatoxin B1) and its use to produce polyclonal antibodies specific to the hapten aflatoxin B-N7-guanine are discussed. An indirect competitive ELISA developed to quantitate aflatoxin adduct showed a 50% inhibition at 15.6 pmol aflatoxin B1-N7-guanine (y=66.73+ (-19.8)x, r=-0.997). Interference by aflatoxin B1 was less than 5%, and no interference by guanine, aflatoxin Gι and aflatoxin M1 was observed. An immunoaffinity column was also developed, by using these polyclonal antibodies, for single-step purification of aflatoxin B1-N7-gua-nine. The immunoaffinity column and the indirect competitive ELISA were evaluated and validated by quantitation of aflatoxin B1-N7-guanine in urine from rats dosed with aflatoxin B1 (1 mg/kg body weight). Spiking studies with standard aflatoxin B1-N7-guanine adduct at 2 and 4 μg/mL phosphate- buffered saline gave 96 and 100% recoveries, respectively, for immunoaffinity cleanup column. The method also was tested successfully for quantitating aflatoxin B1-N7-guanine adduct in spiked human urine. Recoveries of the adduct were 79-90%.

Sign in / Sign up

Export Citation Format

Share Document