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Antibody production wheat

Chongxin Xu, Cunzheng Zhang, Jianfeng Zhong, Hui Hu, Shimin Luo, Xiaoqin Liu, Xiao Zhang, Yuan Liu, Xianjin Liu
In the present study, a Cry1F-immunized rabbit phage display library (6.96×108) was constructed for selecting high activity of anti-Cry1F toxin single-chain antibody (scFv) by bio-panning. Total 16 positive monoclonal phage scFvs were obtained after 4 rounds panning which identified by enzyme-linked immunoassay (ELISA), PCR and DNA sequencing. The most positive phage scFv (named RF4) was expressed in E.coli.HB2151 and the purified soluble protein approximate 30 kDa with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)...
June 16, 2017: Journal of Agricultural and Food Chemistry
Chiara Sacco, Serena Perna, Donatella Vicari, Marco Alfò, Carl-Peter Bauer, Ute Hoffman, Johannes Forster, Fred Zepp, Antje Schuster, Ulrich Wahn, Thomas Keil, Susanne Lau, Paolo Maria Matricardi
BACKGROUND: Previous studies of serum total IgE (t-IgE) were not able to discriminate well enough atopic from non-atopic subjects, i.e. with or without serum-specific IgE antibodies to allergens. OBJECTIVES: To model growth curves of the total IgE levels in children without atopic sensitization (hereafter defined as "normal" t-IgE levels) and to test their usefulness in predicting atopic sensitization. METHODS: The German Multicentre Allergy Study (MAS), a birth cohort with 1314 recruited newborns, began in 1990 and examined the participants until age 20y...
May 25, 2017: Pediatric Allergy and Immunology
Masayuki Morita, Eizo Takashima, Daisuke Ito, Kazutoyo Miura, Amporn Thongkukiatkul, Ababacar Diouf, Rick M Fairhurst, Mahamadou Diakite, Carole A Long, Motomi Torii, Takafumi Tsuboi
The number of malaria vaccine candidates in preclinical and clinical development is limited. To identify novel blood-stage malaria vaccine candidates, we constructed a library of 1,827P. falciparum proteins prepared using the wheat germ cell-free system (WGCFS). Also, a high-throughput AlphaScreen procedure was developed to measure antibody reactivity to the recombinant products. Purified IgGs from residents in malaria endemic areas have shown functional activity against blood-stage parasites as judged by an in vitro parasite Growth Inhibition Assay (GIA)...
April 5, 2017: Scientific Reports
Carmen Gianfrani, Gianfranco Mamone, Barbara la Gatta, Alessandra Camarca, Luigia Di Stasio, Francesco Maurano, Stefania Picascia, Vito Capozzi, Giuseppe Perna, Gianluca Picariello, Aldo Di Luccia
Microwave based treatment (MWT) of wet wheat kernels induced a striking reduction of gluten, up to <20 ppm as determined by R5-antibodybased ELISA, so that wheat could be labeled as gluten-free. In contrast, analysis of gluten peptides by G12 antibody-based ELISA, mass spectrometry-based proteomics and in vitro assay with T cells of celiac subjects, indicated no difference of antigenicity before and after MWT. SDS-PAGE analysis and Raman spectroscopy demonstrated that MWT simply induced conformational modifications, reducing alcohol solubility of gliadins and altering the access of R5-antibody to the gluten epitopes...
March 2017: Food and Chemical Toxicology
S Allahyari-Bake, R Jahanian
Two separate experiments were conducted to investigate the effect of different fat sources and a supplemental exogenous emulsifier (lysophosphatidylcholine, LPC) on growth performance, antibody production titers, and ileal nutrient digestibility in broiler chicks fed with different basal diets. A total of 288 one-day-old Ross 308 chicks were used for each trial (6 dietary treatments based on 3 × 2 factorial arrangements of treatments in both trials) with 4 replicates of 12 birds each. Dietary treatments consisted of 3 different fat sources (soy oil, SO; soy free fatty acids, SFFA; and palm fat powder, PFP) and 2 LPC levels (0 and 0...
May 1, 2017: Poultry Science
Natalia V Beloglazova, Astrid Foubert, Anna Gordienko, Mickael D Tessier, Tangi Aubert, Emile Drijvers, Irina Goryacheva, Zeger Hens, Sarah De Saeger
A sensitive tool for simultaneous quantitative determination of three analytes in one single well of a microtiter plate is shown for the first time. The developed technique is based on use of colloidal quantum dot enrobed into a silica shell (QD@SiO2) derivatives as a highly responsive label. Silica-coated quantum dots were prepared and subsequently modified via the co-hydrolysis with tetraethylorthosilicate (TEOS) and various organosilane reagents. Different surface modification schemes were compared in terms of applicability of the obtained particles for the multiplex immunoassay, e...
November 1, 2016: Talanta
Chongxin Xu, Xiao Zhang, Xiaoqin Liu, Yuan Liu, Xiaodan Hu, Jianfeng Zhong, Cunzheng Zhang, Xianjin Liu
Bt Cry toxin is a kind of bio-toxins that used for genetically modified crops (GMC) transformation widely. In this study, total 15 positive clones could bind the Bt Cry toxins which isolated from a human domain antibody library by 5 rounds affinity selection. According to analyzing of PCR amplification and enzyme-linked immunosorbent assay (ELISA), the most positive phage domain antibody (named F5) gene was cloned into the pET26b vector and expressed in E. coli BL21. The purified antibody was used to develop an indirect competitive ELISA (IC-ELISA) for Cry1Ab, Cry1Ac, Cry1B, Cry1C and Cry1F toxins, respectively...
November 1, 2016: Analytical Biochemistry
Melanie Uhde, Mary Ajamian, Giacomo Caio, Roberto De Giorgio, Alyssa Indart, Peter H Green, Elizabeth C Verna, Umberto Volta, Armin Alaedini
OBJECTIVE: Wheat gluten and related proteins can trigger an autoimmune enteropathy, known as coeliac disease, in people with genetic susceptibility. However, some individuals experience a range of symptoms in response to wheat ingestion, without the characteristic serological or histological evidence of coeliac disease. The aetiology and mechanism of these symptoms are unknown, and no biomarkers have been identified. We aimed to determine if sensitivity to wheat in the absence of coeliac disease is associated with systemic immune activation that may be linked to an enteropathy...
December 2016: Gut
Barbara Lexhaller, Christine Tompos, Katharina Anne Scherf
The safety of gluten-free foods is essential for celiac disease (CD) patients to prevent serious complications. Enzyme-linked immunosorbent assays (ELISAs) are recommended for gluten analysis to monitor the compliance of gluten-free products to the Codex threshold of 20 mg gluten/kg. However, due to the specific features of each gluten ELISA test kit, the results often deviate systematically and largely depend on the characteristics of the antibody. This comprehensive study assessed the specificities and sensitivities of three monoclonal (R5, G12, and Skerritt) and two polyclonal antibodies to the alcohol-soluble prolamin and alcohol-insoluble glutelin fractions of gluten from wheat, rye, and barley, all of which harbor CD-active epitopes...
September 2016: Analytical and Bioanalytical Chemistry
Simon Murch
Recent diagnostic advances have demonstrated that celiac disease is relatively common although most patients have less florid symptoms than previously recognised. The mucosal lesion of this autoimmune disorder depends on both adaptive and innate immune responses. The characteristic antibodies to tissue transglutaminase-2 (tTG-2) and deamidated gliadin peptides may be produced in persons possessing the relevant HLA-DQ genotypes if intact gliadin peptides can penetrate the epithelial barrier to reach antigen presenting cells...
November 2016: Indian Journal of Pediatrics
Yutaro Yamaoka, Shutoku Matsuyama, Shuetsu Fukushi, Satoko Matsunaga, Yuki Matsushima, Hiroyuki Kuroyama, Hirokazu Kimura, Makoto Takeda, Tomoyuki Chimuro, Akihide Ryo
Protein nativity is one of the most critical factors for the quality of antigens used as immunogens and the reactivities of the resultant antibodies. The preparation and purification of native viral antigens in conventional cell-based protein expression systems are often accompanied by technical hardships. These challenges are attributable mainly to protein aggregation and insolubility during expression and purification, as well as to very low expression levels associated with the toxicity of some viral proteins...
2016: Frontiers in Microbiology
Steven C L Ho, Rui Nian, Susanto Woen, Jake Chng, Peiqing Zhang, Yuansheng Yang
Plant and yeast derived hydrolysates are economical and efficient alternative medium supplements to improve mammalian cell culture performance. We supplemented two commercial Chinese hamster ovary (CHO) culture media with hydrolysates from four different sources, yeast, soybean, Ex-Cell CD (a chemically defined hydrolysate replacement) and wheat to improve the productivity of two cell lines expressing different monoclonal antibodies (mAbs). Yeast, soybean and Ex-Cell CD improved the final mAb titer by increasing the specific productivity (qP) and/or extension of the culture period...
October 2016: Journal of Bioscience and Bioengineering
Zhaowei Zhang, Xiaofeng Hu, Qi Zhang, Peiwu Li
Mycotoxins usually found in agricultural products such as peanut, corn, and wheat, are a serious threat to human health and their detection requires multiplexed and sensitive analysis methods. Herein, a simultaneous determination for aflatoxin B1, B2, G1, G2, ochratoxin A, zearalanone and T-2 toxin was investigated using high performance liquid chromatography coupled with tandem mass spectrometry in a single run via a home-made multiple immunoaffinity column. Four monoclonal antibodies were produced in our lab against aflatoxins, ochratoxin A, zearalanone and T-2 toxin, respectively, then combined as a pool and bound to Sepharose-4B for affinity chromatography...
May 15, 2016: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Kayoko Matsunaga, Yasutaka Kuroda, Shinobu Sakai, Reiko Adachi, Reiko Teshima, Akiko Yagami, Hiroshi Itagaki
Recent reports suggest that hydrolyzed wheat protein (HWP) variants such as Glupearl® 19S (GP19S) induce immediate-type hypersensitivity via epicutaneous (EC) sensitization. The identification of strong allergens is a key step in product assessment before commercial launch. However, few reports have described the estimation of actual and potential anaphylactic sensitizing capacity. In this study we assessed the strength of both the actual and potential anaphylactic sensitizing capacity by investigating the immediate-type hypersensitivity inducing potential of HWP compared with gluten...
December 2015: Journal of Toxicological Sciences
K A Scherf, K Brockow, T Biedermann, P Koehler, H Wieser
Wheat-dependent exercise-induced anaphylaxis (WDEIA) is a rare, but potentially severe food allergy exclusively occurring when wheat ingestion is accompanied by augmenting cofactors. It is clinically characterized by anaphylactic reactions ranging from urticaria and angioedema to dyspnoea, hypotension, collapse, and shock. WDEIA usually develops after ingestion of wheat products followed by physical exercise. Other cofactors are acetylsalicylic acid and other non-steroidal anti-inflammatory drugs, alcohol, and infections...
January 2016: Clinical and Experimental Allergy: Journal of the British Society for Allergy and Clinical Immunology
Azadeh Niknejad, Diane Webster, Mrinal Bhave
The emergence of antibiotic resistant pathogenic strains of bacteria has necessitated the development of novel antimicrobial agents. The puroindoline A and B (PINA and PINB) proteins of wheat, well-known for their roles in determining the important phenotype of grain texture, are also antimicrobial, making them attractive as natural bio-control agents. However, the biochemical basis of PIN functionality remains unclear due to limitations in expressing them at the required yield and purity and lack of accurate tertiary structure...
September 2016: Protein Expression and Purification
Hélène Garnier-Lengliné, Nadine Cerf-Bensussan, Frank M Ruemmele
Celiac disease is an autoimmune enteropathy, triggered by ingestion of gluten in genetically predisposed individuals. Since the use of anti-transglutaminase and anti-endomysium antibodies in the early 1990s, two main groups of clinical presentation can be identified: patients with a symptomatic form of the disease, and patients with a pauci (a)-symptomatic form detected during the work-up of another autoimmune disease or due to a family history of celiac disease. The prevalence of both forms of the disease is currently estimated between 1/100 and 1/400...
October 2015: Clinics and Research in Hepatology and Gastroenterology
Naiyana Gujral, Ju Won Suh, Hoon H Sunwoo
BACKGROUND: Pepsin-trypsin resistant gliadin (PT-gliadin) promotes intestinal tissue inflammation and increases paracellular permeability of immunogenic gliadin peptides into the lamina propria. This leads to the complications seen in the pathogenesis of celiac disease (CD). In this study, specific anti-gliadin IgY antibody was produced and evaluated for its efficacy on gliadin induced intestinal integrity impairment and proinflammatory effects on intestinal epithelial (Caco-2) cell culture model for CD...
July 9, 2015: BMC Immunology
Sachiko Onishi, Endy Adnan, Jun Ishizaki, Tatsuhiko Miyazaki, Yuki Tanaka, Takuya Matsumoto, Koichiro Suemori, Masachika Shudou, Takafumi Okura, Hiroyuki Takeda, Tatsuya Sawasaki, Masaki Yasukawa, Hitoshi Hasegawa
Systemic lupus erythematosus (SLE) is characterized by production of a variety of autoantibodies. Although anti-double-stranded DNA (anti-dsDNA) antibodies contribute to the pathogenesis of lupus nephritis (LN), they are not sufficient for diagnosis and evaluation of disease activity. To obtain other autoantibodies associated with LN, we screened autoantigens reacting with the sera of LN patients by using an N-terminal biotinylated protein library created from a wheat cell-free protein production system. We screened 17 proteins that showed higher positive signals in the active phase than in the inactive phase of SLE, and higher positive signals in the serum of SLE patient with nephritis than in that of patient without nephritis...
2015: PloS One
Bharani Srinivasan, Margarete Focke-Tejkl, Milena Weber, Sandra Pahr, Alexandra Baar, Raja Atreya, Markus F Neurath, Harald Vogelsang, Wolf-Dietrich Huber, Rudolf Valenta
BACKGROUND: Celiac disease (CD) is an inflammatory disease of the small intestine caused by an immunologic hypersensitivity reaction to dietary wheat gluten. OBJECTIVES: We sought to clone, express, and perform IgA epitope mapping of a CD-specific wheat antigen and to study its usefulness for identifying patients with CD and monitoring adherence to a gluten-free diet. METHODS: A synthetic gene coding for γ-gliadin 1 (GG1) was expressed in Escherichia coli...
December 2015: Journal of Allergy and Clinical Immunology
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