Authors:Q. Li, K. Ji, X. Song, E. Zhang, Y. Pei, and Y. Yan
Gliadins were extracted from three wheat varieties, viz. Nongda 99260037 (good quality), Henan 9023 (media quality) and Nongda 98123 (poor quality), and
α-, β-, γ-, ω
-gliadins were purified from Nongda 99260037 using preparative SDS-PAGE. The total gliadins and
α-, β-, γ-, ω
-gliadins were used as antigens to immunise BALB/C mice, and the corresponding polyclonal antibodies were prepared, designated as Anti-A, Anti-B, Anti-C, Anti-A
, respectively. Binding of the polyclonal antibodies with 8 varieties, that varied in quality properties, showed correlations with some wheat quality parameters. Correlation coefficients between antibodies against total gliadins or
-gliadin of Nongda 99260037 and quality parameters were higher than other antibodies. Of seven polyclonal antibodies tested, three (Anti-A
and Anti-A) displayed significant positive or negative associations between antibody binding and dough development time, strength, valorimenter value and stability time, but no significant correlations were observed with water absorption. These results suggest that polyclonal antibodies could be used for rapid prediction and screening of wheat quality parameters.
Authors:Kh. Sallam, I. Abdel-Ghany, H. Abdel-Aziz, and T. Siyam
The aim of this work is the development of the immunogenic properties of 17beta-estradiol to produce polyclonal antibody for
radioimmunoassay purposes. Poly(acrylamide–maleic acid) gel [P(AAm–MA)] was prepared by the template polymerization. Polyacrylamide
(PAAm) was used as a template for the polymerization of MA in aqueous solution using gamma rays as initiator. Then a comparative
study of estradiol trapped in the prepared polymer gels versus estradiol conjugated to bovine serum albumin (BSA) was carried
out. Ten rats divided into two groups were injected according to specific protocol. The immunogenic properties of both produced
polyclonal antibodies were studied. In conclusion, the results revealed that the similar immunogenic properties with specific
radiolabeled tracer were obtained.
Authors:S. Milesz, Yu. Norseev, Z. Szücs, and L. Vasáros
The complex formation of astatine/I/ cation with diethylene triamine pentaacetic acid /DTPA/ and characterization of the complexes were investigated by electromigration in free electrolytes and by gel-chromatography on Sephadex G 25. We describe the conjugation procedure for the production of At-DTPA conjugated polyclonal antibodies.
Polyclonal immunoglobulin IgG was labeled using three different methods: a direct method via 2-mercaptoethanol, an indirect
one, in which a chelating group was covalently attached to the protein and the99mTc added as a glucoheptonate complex and a photoactivation method. The properties of99mTc polyclonal antibody labeled by three different methods were assessed by in vitro and in vivo studies. The ratio between
inflamed thigh to normal thigh was similar and independent of the method of labeling.
The purpose of this work was to label polyclonal antibodies with99mTc such as photoactivated IgG and to check the radiochemical and biological behavior of the labeled product. Experiments were
carried out to found the formulation of optimal binding of99mTc to polyclonal IgG. In addition animal studies in normal mice and in mice bearing a promoted inflammation foci were performed.
The labeled product was analyzed by size-exclusion HPLC and ITLC. Higher amount than 23μg of tin per 500μg of protein gave
between 87% and 93% labeling. Protein concentrations between 1.5 and 5 mg/ml gave 90% labeling yields.
Authors:H. Shafik, N. El-Mouhty, A. EL-Bayoumy, and I. Abdel-Ghany
Polyclonal antibody production in mammals is generally associated with multiple injections of antigens and adjuvant and repeated
blood sampling procedures. In the production of second polyclonal antibodies, a number of critical steps can be identified
that may influence the outcome of the animal experiment (immunological results and the pain and suffering of the animals).
The goal of this work was to evaluate critical steps in the production of these antibodies, and to optimize production protocols
that will ultimately result in effective antibody responses. This work was achieved through immunization of two healthy sheep
by purified Alpha feto protein (AFP) antigen. Also, the present study involved the preparation of AFP standards from human
cord blood. Furthermore, preparation of a radiolabeled AFP tracer of a high specific activity using 125I isotope by chloramine T method was undertaken. Moreover, this study provides that there was no observable difference between
the Scottish Antibody Production Unit (SAPU) and Sigma SAM IgG and the two second antibodies obtained from the local sheep
Porcine epidemic diarrhoea virus (PEDV) is one of the important pathogens that may cause severe diarrhoea in piglets. In this study, the nucleocapsid (N) gene of a Chinese PEDV isolate designated HLJBY was cloned. The phylogeny of PEDV strains was investigated by constructing a phylogenetic tree based on the N protein sequences. The results indicate that there are two major groups of Chinese PEDVs, a Japanese PEDV group and a Korean PEDV group. High-level expression of the N protein was achieved in Escherichia coli. The immunoreactivity between PEDV particles or the bacterially expressed N protein and rabbit anti-PEDV serum was confirmed by immunofluorescence assays and Western blot. Both PEDV N protein and the polyclonal antibody generated in this study are valuable diagnostic reagents for PEDV surveillance.
The distribution of EGF receptors (EGF-R) was examined in normal, hyaline membrane diseased and pneumonic newborn lung tissues by immunohistochemical methods under the light microscope. The PAP technique with polyclonal antibodies was performed to demonstrate the EGF receptor localisation in these tissues. Strong EGF-R reactivity was observed on bronchiolar epithelium and type I and type II alveolar cells in normal newborn loung tissues; whereas, poor reactivity was observed in alveolar macrophages. On the other hand, strong immunoreactivity was detected in type I alveolar cells and alveolar macrophages in hyaline membrane disease, but no reactivity was present in type II alveolar cells. The strongest immunoreactivity was observed in alveolar macrophages of newborn pneumonic lung tissues. In conclusion, the most meaningful form of reactivity was observed in normal newborn lung tissues of airway track and respiration area. This result is related with the maturation of the lungs after birth.
Authors:B. Mickowska, P. Socha, D. Urminská, and E. Cieślik
Prolamins are alcohol-soluble fraction of cereal proteins involved in immunological response of patients with celiac disease. The aim of this study was to analyse the similar protein complex of selected varieties of cereal, pseudocereal and legume grains by comparison of protein fractions, amino acids composition and SDS-PAGE electrophoresis. The immunoreactivity was tested by Western blot and ELISA methods. ELISA analysis recognizes celiac active epitopes in wheat gliadin (which is reference protein in determination of celiac activity), and also corresponding epitopes in other grain proteins responsible for immunological response of patients with celiac disease. Estimated quantity of celiac active sequences (calculated as gliadin content) below 20 ppm was found in species of amaranth, buckwheat and millet, as well as rice, maize, chickpea and chickling vetch. Immunological reaction with polyclonal antibody was negative for all crops, except oat, maize, millet and foxtail millet.