• Title/Summary/Keyword: Immunoglobulin G.

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Production of Egg Yolk Immunoglobulin and Its Application for Pepper mild mottle virus in Serological Tests (Pepper mild mottle virus에 대한 난황항체의 생산과 혈청학적 진단에의 활용)

  • Han Jung-Heon;Lee Cheol-Ho;Kim Young-Ho;La Yong-Joon
    • Research in Plant Disease
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    • v.12 no.2
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    • pp.144-147
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    • 2006
  • Egg yolk immunoglobulin (IgY) is much widely used in medical fields, but its use in serology of plant viruses is much limited. We produced an IgY against pepper mild mottle virus (PMMoV) and applied it to several serological tests. Polyclonal antibodies were obtained from the egg yolk of chicken immunized with a total of 2mg of purified PMMoV over 2 months. The titers of antibodies were measured with the ring-test over six months after the first injection. The highest.titers of IgY was 1/2,560 at 2 months after the first injection. Approximately 60-80 mg of IgY were obtained from one egg yolk. Using the IgY, 1ng/ml of purified PMMoV was detected with the indirect ELISA. Gelrite gel double diffusion test, ELISA and tissue immuno-binding assay employing IgY gave similar sensitivity and specificity to those of IgG developed in rabbit. Therefore, the IgY which can be obtained in large quantities from a chicken, might be useful for the antibody production and the serology of plant viruses.

Detection of IgG and IgM antibodies with ELISA technique in human trichomoniasis (질트리코모나스 환자에서 효소표식 면역검사법을 이용한 혈청 내 항-질트리코모나스 IgG 및 IgM 항체가의 측정)

  • 이미리;신명헌
    • Parasites, Hosts and Diseases
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    • v.28 no.1
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    • pp.25-30
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    • 1990
  • The direct wet mount examination of vaginal. secretion, widely applied for the diagnosis of Trichcmonas vaginalis infection in woman patients, is rapi4 and economical, however, the sensitivity of this technique is not so high. In this study enzyme-linked immunosorbent assay (ELISA) was employed for the detection of serum anti-T. vaginalis IgG and IsM antibodies from 30 vaginal trichomoniasis patients and 30 non-infected healthy persons. The results were as follows: 1. Serum ELISA-IgG value was $0.37{\pm}0.134(Mean{\pm}S.D.)$ in vaginal trichomoniasis patients and $0.21{\pm}0.054$ in healthy controls(P<0.005), and the sensitivity and specificity of ELISA for serum IgG antibody were 70.0% and 96.7%, respectively. 2. Serum ELISA-IgM value was $0.33{\pm}0.177 (Mean{\pm}S.D.)$ in vaginal trichomonlasls patients and $0.11{\pm}0.051$ in healthy controls (p<0.005), and the sensitivity and specificity of ELISA for serum IsM antibody were 70.0% and 96.7%, respectively. 3, The ELISA-IgG values showed a significant correlation with ELISA-IgM values(r=0.77, p<0.005) , With above results, it is assumed that ELISA is a reliable method for the diagnosis of T vaginalis infection and simultaneous measurement of serum IgG and IgM with this technique is recommended.

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Comparison of TIA with ELISA for circulating antibody detection in clonorchiasis (간흡충증에 있어서 항체검출을 위한 Enzyme-linked Immunosorbent Assay와 Thin Layer Immunoassay의 비교)

  • 이용기;유재숙이근태정경일
    • Parasites, Hosts and Diseases
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    • v.21 no.2
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    • pp.265-269
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    • 1983
  • A comparison was made of a new serological method, thin layer immunoassay (TIA), and an established method, enzyme-linked immunosorbent assay (ELISA), in the detection and quantiacation of antibodies in clonorchiasis. Saline extract of Iyophilized Clonorchis sinensis adult worm was used as antigen, and TIA by the method of Elwing et at. (1976) and ELISA by Voller et at. (1974) were performed. Using sera from known clonorchiasis cases,100% of the sera tested were Positive by TIA and 88.35 by ELISA. TIA produced false positive results in 14 out of 36 cases, which were 10 amoebiasis cases, 16 paragonimiasis cases and 10 healthy controls. ELISA. however, produced a small number of false positives, 7 out of 55 cases. There was correlation between Immunoglobulin G level in sera and ELISA value (correlation coefficient, 0.69), whereas no correlation between Immunoglobulin G level and TIA result. The Performance of TIA and ELISA was not correlated in the results using homologous antigen.

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Interaction between Whey and Soybean Proteins (유청 및 대두 단백질의 상호작용)

  • Shon, Dong-Hwa;Lee, Hyong-Joo
    • Applied Biological Chemistry
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    • v.31 no.4
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    • pp.361-370
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    • 1988
  • To investigate the interaction between whey and soybean protein, thermal changes of component proteins were analyzed by column chromatography and gel electrophoresis. In the Sephadex G-200 chromatography of the mixture treated at above $80^{\circ}C$, the amount of low molecular weight proteins and high molecular aggregates were increased. This implicated that dissociation of 1ls globulin into subunits and the formation of soluble aggregates between these subunits and whey proteins that contain thiol and disulfide groups. These interaction between soy proteins and ${\beta}-lactoglobulin$, ${\alpha}-lactalbumin$, and proteose-peptone 3 were confirmed by gel electrophoresis. Bovine serum albumin, Immunoglobulin-G(H), Lactoferrin, 1ls-subunits(basic and acidic), and subunit of 7s globulin were also considered to interact each other depending on the condition of the salt solutions.

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AN IMMUNOHISTOCHEMICAL STUDY ON THE IMMUNOGLOBULINS OF EXPERIMENTALLY INDUCED RAT PERIAPICAL LESIONS (실험적 백서 치근단 병소에서의 면역글로불린 분포에 관한 면역조직화학적 연구)

  • Boo, Jung-Sun;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.15 no.2
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    • pp.58-76
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    • 1990
  • This study was designed to elucidate the distribution of the immunoglobulins in the experimentally induced rat periapical lesions. The pulp exposure was performed in 80 molars from 40 rats and the animals were sacrificed at 15, 30, 60 and 90 days after the operation and examined and radiographed. Of the 80 samples, 56 samples were routinely sectioned ($4-6{\mu}$ in thickness) and stained with Hematoxylin-Eosin for the light microscopic examination and 50 samples were stained with toluidin blue for mast cells and 50 samples were stained using the Avidin-Biotin horseradish peroxidase for detecting the presence of Ig A, Ig E, Ig M and Ig G containing cells. The following results were obtained : 1. The periapical lesions could be observed in all of 80 teeth by radiogragh (100%) and the periapical lesions were detected in 50 samples of 51 samples by light microscopy (98%). The size of lesions increased with time lapse both by radiograph and by light microscopy(p<0.05). 2. Of the 50 samples, 19 samples were diagnosed as periapical abscesses, 18 as periapical granulomas, 10 as fibrous scar tissues and 3 cysts. 3. After pulp exposure, periapical granulomas were developed mostly in the 15 day group, with time lapse periapical abscesses and fibrous scar tissues increased. 4. In the 50 periapical lesions, the numbers of Ig G containing cell (57.2%) were prominent and the percentage of Ig A, Ig E and Ig M containing cells were 16.4%, 14.7% and 11.8% respectively. The numbers of all classes of immunoglobulin containing cell were highest in the periapical granulomas and lowest in the cysts(p<0.05). 5. The number of the mast cell and immunoglobulin containing cells decreased generally with time lapse after the pulp exposure and Ig A, Ig E, Ig M and Ig G containing cells and mast cells had the high correlation one another(>0.6).

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Protective efficacy of a novel multivalent vaccine in the prevention of diarrhea induced by enterotoxigenic Escherichia coli in a murine model

  • Zhao, Hong;Xu, Yongping;Li, Gen;Liu, Xin;Li, Xiaoyu;Wang, Lili
    • Journal of Veterinary Science
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    • v.23 no.1
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    • pp.7.1-7.14
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    • 2022
  • Background: Enterotoxigenic Escherichia coli (ETEC) infection is a primary cause of livestock diarrhea. Therefore, effective vaccines are needed to reduce the incidence of ETEC infection. Objectives: Our study aimed to develop a multivalent ETEC vaccine targeting major virulence factors of ETEC, including enterotoxins and fimbriae. Methods: SLS (STa-LTB-STb) recombinant enterotoxin and fimbriae proteins (F4, F5, F6, F18, and F41) were prepared to develop a multivalent vaccine. A total of 65 mice were immunized subcutaneously by vaccines and phosphate-buffered saline (PBS). The levels of specific immunoglobulin G (IgG) and pro-inflammatory cytokines were determined at 0, 7, 14 and 21 days post-vaccination (dpv). A challenge test with a lethal dose of ETEC was performed, and the survival rate of the mice in each group was recorded. Feces and intestine washes were collected to measure the concentrations of secretory immunoglobulin A (sIgA). Results: Anti-SLS and anti-fimbriae-specific IgG in serums of antigen-vaccinated mice were significantly higher than those of the control group. Immunization with the SLS enterotoxin and multivalent vaccine increased interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations. Compared to diarrheal symptoms and 100% death of mice in the control group, mice inoculated with the multivalent vaccine showed an 80% survival rate without any symptom of diarrhea, while SLS and fimbriae vaccinated groups showed 60 and 70% survival rates, respectively. Conclusions: Both SLS and fimbriae proteins can serve as vaccine antigens, and the combination of these two antigens can elicit stronger immune responses. The results suggest that the multivalent vaccine can be successfully used for preventing ETEC in important livestock.

Tandem Mass Spectrometry of N-linked Glycans from Human Immunoglobulin G (다중 질량 분석법을 이용한 인체 면역글로불린 G의 N-연결 글라이칸 분석)

  • Joo, Hwang-Soo;Kim, Yun-Gon;Jang, Kyoung-Soon;Kim, Byung-Gee
    • KSBB Journal
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    • v.22 no.4
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    • pp.234-238
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    • 2007
  • We used electrospary ionization ion trap tandem mass spectrometry (ESI-IT tandem MS) to structural elucidation of three different biantennary-type glycans having zero, one, two galactoses (G0, G1, G2). The highest fragment ion in the MS/MS spectra of three glycans was produced by 0,2-ring cleavage of fucose-linked N-acetylglucosamine (GlcNAc) in reducing end. The fragment ions both from precursor ions and 0,2-ring cleaved ions ($^{0.2}An$; n=5 for G0, n=6 for G1 and G2) were not overlapped each other. As results of $MS^n$ analyses, tandem fragmentation trees of each glycans were generated and 2,4-ring cleavages ($^{2.4}A_6$) were occurred in GlcNAc linked to reducing end GlcNAc. This structural elucidation and fragmentation study of N-linked glycans by tandem mass spectrometry can be applied to structural analysis of more complicated glycans.

Does supplementing laying hen diets with a herb mixture mitigate the negative impacts of excessive inclusion of extruded flaxseed?

  • Hossein Hosseini;Noah Esmaeili;Aref Sepehr;Mahyar Zare;Artur Rombenso;Raied Badierah;Elrashdy M. Redwan
    • Animal Bioscience
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    • v.36 no.4
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    • pp.629-641
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    • 2023
  • Objective: This study investigated the effects of extruded flaxseed with and without herbs mixture on egg performance, yolk fatty acids (FAs), lipid components, blood biochemistry, serological enzymes, antioxidants, and immune system of Hy-Line W-36 hens for nine weeks. Methods: Two hundred forty laying hens were randomly distributed to eight treatments, resulting in six replicates with five hens. Graded levels of dietary extruded flaxseed (0, 90, 180, and 270 g/kg) with and without herbs mixture (24 g/kg: garlic, ginger, green tea, and turmeric 6 g/kg each) were designed as treatments. Results: The two-way analysis of variance indicated that hens fed herbs mixture had a higher value of egg production, yolk high-density lipoprotein (HDL), superoxide dismutase, glutathione peroxidase, and white blood cell and lower contents of yolk cholesterol, glucose, and blood low-density lipoprotein than those fed diets without herb mixtures (p<0.05). The Flx27 (270 g/kg flaxseed) (153.5 g/kg n-3 FAs) and Flx27+H (270 g/kg flaxseed plus 24 g/kg herbs mixture) (150.5 g/kg n-3 FAs) groups were the most promising treatments in terms of yolk n-3 FAs content. In-teraction effect (herbs- flaxseed) for blood cholesterol, HDL, malondialdehyde, glutaredoxin, alanine transaminase, (ALT), aspartate transaminase (AST), haemoglobin and immune parameters was significant (p<0.05). The results showed layers fed herbs mixture (Flx9+H, Flx18+H, and Flx27+H) had a better value of total antibody, immunoglobulin M, immunoglobulin G, ALT, AST, and blood HDL as compared with representative flaxseed levels without herbs. Conclusion: High inclusion levels of extruded flaxseed (270 g/kg) without herbs to enrich eggs with n-3 appears to impair the antioxidant system, immunohematological parameters, and sero-logical enzymes. Interestingly, the herbs mixture supplementation corrected those effects. Therefore, feeding layers with flaxseed-rich diets (270 g/kg) and herbs mixture can be a promising strategy to enrich eggs with n-3 FAs.

Fabrication of Protein A-Viologen Hetero LB Film for Antibody Immobilization

  • Lee, Heon-Ju;Choe, Jeong-U;Lee, U-Chang;O, Byeong-Geun;Lee, Won-Hong
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.859-862
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    • 2001
  • For the development of preferable immunosensor and protein chip, the viologen Langmuir-Blodgett (LB) multilayer was fabricated on the surface, and then protein A was adsorbed on the proposed viologen LB film by electrostatic attractive force. The Immunoglobulin G (IgG) labeled with fluorescence marker was self-assembled on the fabricated protein A film. The topographies of the deposited films were investigated by using atomic force microscope (AFM). The immobilization of IgG was verified by fluorescence spectrum. Such structures can be used as sublayers for various kinds of IgG immobilization toward immunosensors and protein chip.

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Partial characterization of a 29kDa cysteine protease purified from Taenia solium metacestodes

  • KIM Ji-Young;YANG Hyun-Jong;KIM Kwang-Sig;CHUNG Young-Bae
    • Parasites, Hosts and Diseases
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    • v.43 no.4 s.136
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    • pp.157-160
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    • 2005
  • A 29kDa cysteine protease of Taenia solium metacestodes was purified by Mono Q anion-exchanger and Superose 6 HR gel filtration chromatography. The enzyme was effectively inhibited by cysteine protease inhibitors, such as iodoacetic acid (IAA) and trans-epoxy-succinyl-L-leucyl-amido (4-guanidino) butane (E-64) while inhibitors acting on serine- or metallo-proteases did not affect the enzyme activity. The purified enzyme degraded human immunoglobulin G (IgG), collagen and bovine serum albumin (BSA), but human IgG was more susceptible for proteolysis by the enzyme. To define the precise biological roles of the enzyme, more detailed biochemical and functional studies would be required.