• Title/Summary/Keyword: serum IgG antibody

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Purification of IgG1 Type Mouse Monoclonal Antibodies with DEAE-Trisacryl Chromatography (DEAE-Trisacryl 크로마토그래피법에 의한 IgG1 Type 쥐 단일클론 항체의 분리정제)

  • 최태부;정용근
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.335-342
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    • 1988
  • An anion exchange chromatography was employed for the purification of mouse monoclonal antibodies from ascitic fluid and in vitro cultivation media. After cultivation of hybridomas, Alps 25-3, HCGK, A4W, and KW, producing IgG1, the culture supernatants were harvested by centrifugation, precipitated with 50-60% ammonium sulfate, and dialyzed against 0.025 M Tris-HCI buffer (pH 8.2). Then the dialyzed samples were loaded into a DEAE-Trisacryl M anion exchange column. Monoclonal antibodies bound to the DEAE-Trisacryl M were eluted with 0.025 M Tris-HCI buffer (pH 8.2) containing 30-40 mM NaCl. In ammonium sulfate precipitation, the recovery of the monoclonal antibody was shown to be 90% and 84% from in vitro culture media containing 10% and 2% fetal bovine serum, respectively. On the other hand, the pretreatment by ultrafiltration enhanced the yield up to 91% whereas the purity was lower than that by ammonium sulfate treatment. Subsequently, in the DEAE-Trisacryl M chromatographic separation, the purities and recoveries of all the monoclonal antibodies from both the in vitro culture supernatants and ascitic fluids were 70-80% and 65% respectively. The monoclonal antibody, Alps 25-3 could be further purified with a purity of 95% through an immunoadsorbent chromatography.

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Effect of golden needle mushroom (Flammulina velutipes) stem waste on laying performance, calcium utilization, immune response and serum immunity at early phase of production

  • Mahfuz, Shad;Song, Hui;Liu, Zhongjun;Liu, Xinyu;Diao, Zipeng;Ren, Guihong;Guo, Zhixin;Cui, Yan
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.5
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    • pp.705-711
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    • 2018
  • Objective: This experiment was conducted to evaluate the effects of golden needle mushroom (Flammulina velutipes) stem waste (FVW), on organic eggs production, calcium utilization, antibody response, serum immunoglobulin, and serum cytokine concentration at early phase of production in laying hens. Methods: A total 210, 19 weeks old aged ISA Brown layers were randomly assigned into 5 equal treatment groups, with 7 replications of 6 hens each. Dietary treatment included a standard basal diet as control; antibiotic (0.05% flavomycin); 2% FVW; 4% FVW; and 6% FVW. The experimental duration was 10 weeks. Results: There was no significant differences (p>0.05) on hen day egg production, egg weight, egg mass, feed intake, and feed conversion ratio (FCR) among experimental groups. Unmarketable eggs were significantly lower (p<0.05) both in 4% FVW and 6% FVW fed groups than control group. The calcium retention and calcium in egg shell deposition were significantly higher (p<0.05) in FVW inclusion groups than control and antibiotic groups. Antibody titers against Newcastle diseases were significantly higher (p<0.05) in 6% FVW fed group (except combined with 4% FVW at day 147) and infectious bronchitis were significantly higher (p<0.05) in FVW fed groups (except 2% FVW and 4% FVW at day 161) than control and antibiotic groups. Serum immunoglobulin sIgA was significantly higher (p<0.05) in all levels of FVW and IgG was significantly higher (p<0.05) in 4% FVW than control and antibiotic groups. Serum cytokine concentration interleukin-2 (IL-2) was significantly higher (p<0.05) in 6% FVW; IL-6 and tumor necrotic $factor-{\alpha}$ were significantly higher (p<0.05) both in 4% FVW and 6% FVW than control and antibiotic groups; IL-4 was significantly higher (p<0.05) in antibiotic, 2% FVW and 4% FVW fed groups than control. Conclusion: F. velutipes mushroom waste can be used as a novel substitute for antibiotic for organic egg production and sound health status in laying hens.

The effect of kaolin and acetone-ether treatment on HI antibody titres to arboviruses in chicken serum (Kaolin 및 Acetone-ether 처리가 항뇌염병독(抗腦炎病毒) 계혈청(鷄血淸)의 특이 항체에 미치는 영향)

  • Chung, Y.S.
    • Korean Journal of Veterinary Research
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    • v.9 no.2
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    • pp.1-5
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    • 1969
  • 닭 혈청(血淸)속에 존재(存在)하는 뇌염병독(腦炎病毒)에 대(對)한 비특이성(非特異性) 억제 물질(物質)을 제거(除去)하기 위하여 Kaolin 또는 Acetone-ether로서 그 혈청(血淸)을 처리 하였을때 그 처리가 비특이성(非特異性) 억제물질(物質) 뿐만 아니라 특이(特異)한 항체(抗體)의 역가(力價)에 대(對)한 영향의 유무(有無)를 시험하였다. Kaolin 및 Acetone-ether 처리는 확실히 특이항체(特異抗體)의 HI 역가(力價)의 감소(減少)를 가져왔다. IgM type 항체는 Kaolin 처리에 의해서 그리고 IgG type 항체는 Acetone-ether 처리에 의해서 더 큰 항체역가(抗體力價)의 감소(減少)를 가져 왔다.

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Assignment of the Carbonyl Carbon Resonances in Anti-Dansyl Antibodies (항 단실 항체의 카르보닐탄소 유래 시그날의 귀속)

  • ;;Koichi Kato;Yoji Arata
    • YAKHAK HOEJI
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    • v.39 no.5
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    • pp.516-520
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    • 1995
  • The anti-dansyl antibodies were specifically labeled with stable isotope by growing hybridoma cells in serum-free medium. Assignments of the observed carbonyl carbon resonances have been determined by using $^{13}C-{15}N$ double labeling method in order to assign the Leu resonances. However, when the identical dipeptide appears more than twice in the polypeptide sequences, we applied the proteolytic fragments in the fragment-specific method. Carboxypep-tidase B-treated antibody has also been used to assign the Lys-447 in C terminal amino acid. These unambiguously assigned carbonyl carbon resonances in antibodies are thought to be useful in elucidating not only the structure of antibodies but also the structure-function relationship in the antibody by $^{13}C$ neuclear magnetic resonance spectroscopy.

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ANTIBODY PRODUCTION BY PARENTERAL ADMINISTRATION OF STREPTOCOCCUS MUTANS AND GLUCOSYLTRANSFERASE IN MICE (비경구 투여한 Streptococcus mutans 균체 및 Glucosyltransferase에 대한 마우스의 면역항체반응)

  • Yang, Kyu-Ho;Chung, Mee;Chung, Jin;Chang, Mee-Young;Oh, Jong-Suk;Nah, Hee-Sam;Kang, In-Chol;Lee, Hyun-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.1
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    • pp.61-68
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    • 2003
  • Streptococcus mutans is known to be a major causative organism of human dental caries. The development of a vaccine against dental caries involves identification of appropriate antigens of mutans streptococci against which protective immune responses can be mounted, and the selection of a method of immunization that will generate sustained levels of protective antibodies. Antigens receiving most attention include streptococcal surface proteins that are involved in attachment to tooth surfaces and glucosyltransferases (GTF) that synthesize adhesive glucans from sucrose. The induction of antibody responses to orally administered antigens is often difficult due to digestive destruction of antigens and immune tolerance. Here we report the induction of antibody responses to an anti-caries vaccine containing retinoic acid (RA). Subcutaneous immunization with formalin-fixed bacteria or GTF supplemented with RA induced higher serum IgM and IgA responses to GTF compaired to oral adminstration. Antisera induced by Ingbritt strain showed partial cross-reaction with LM-7 strain, but not with OMZ175. These results suggest that subcutaneous immunization with GTF combined with an immunomodulator, RA, may be applied to anti-caries vaccine.

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Effect of Antisera from Clostridium difficile-Infected Mice on Toxin-A-Induced Colonic Epithelial Cell Death Signaling

  • Kim, Dae Hong;Lee, Ik Hwan;Nam, Seung Taek;Nam, Hyo Jung;Kang, Jin Ku;Seok, Heon;Hwang, Jae Sam;Kim, Ho
    • Journal of Microbiology and Biotechnology
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    • v.24 no.5
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    • pp.696-703
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    • 2014
  • Clostridium difficile causes mucosal damage and diarrhea by releasing two exotoxins: toxin A and toxin B. C. difficile colitis is associated with alterations in bowel flora and the failure to mount an effective antibody response. The aim of the current study was to investigate whether antitoxin sera prevent toxin-A-induced apoptosis, cytoskeletal disaggregation, cell detachment, and tight junction loss in cultured colonic epithelial cells. Serum samples were isolated from mice that survived a C. difficile infection following antibiotic treatment, and the antitoxin effects of these samples were investigated in toxin-A-exposed HT29 colonic epithelial cells and a toxin-A-induced animal model of gut inflammation. Unchallenged mice did not produce IgG against toxin A, whereas serum (antiserum) from C. difficile-challenged mice showed significant IgG responses against toxin A. Treatment with the antiserum markedly inhibited mucosal damage and inflammation in the toxin-A-treated mouse model. In contrast to control mouse serum, the antiserum also markedly inhibited toxin-A-induced DNA fragmentation, dephosphorylation of paxillin and Epo receptor (EpoR), deacetylation of tubulin, and upregulation of p21(WAF1/CIP1) and p53. Taken together, these results reveal that the generated antitoxin serum has biotherapeutic effects in preventing various C. difficile toxin-A-induced cellular toxicities.

A Study of Korean Dietary Lipid Sources on Lipid Metabolism and Immune Function in Rat (한국인 상용식이지방이 흰쥐의 지방대사 및 면역능력에 미치는 영향)

  • 이종미
    • Journal of Nutrition and Health
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    • v.20 no.5
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    • pp.350-366
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    • 1987
  • This research was designed to study the effect of Korean dietary lipids on the lipid metabolism and the immune function in young rats . The three different lipid sources were compared lard, perilla oil and fish oil. Three different levels of lipid in the diet, 2%, 15% and 30%,on the weight basis, were included. After four weeks feeding, the rats were sacrified and blood sample was collected to analyze for the total lipid, TG and cholesterol contents in serum. The HDL fraction in serum was seperated by the electrophoresis of lipoproteins. The immune responses were measured by the blastogenesis of spleen lymphocyte stimulated by PHA and in serum were measured. The following results were obtained. Lower body weight gain was shown in 30% lipid diet fed group on the isocaloric basis. In concerning the different dietary lipid sources, there were significantly lower boyd weight gain in fish oil than in perilla seed oil and lard group in 30% lipid groups. Deposition of body fat expressed by epididymal fat pad in serum were significantly different among perilla seed oil, lard and fish oil groups. Perilla seed oil group showed lowest level of total lipid and TG in serum regardless of dietary fat level. The feeding perilla seed oil to rats was resulted in lower serum cholesterol levels than lard in all three levels of fats tested. The HDL fraction was elevated in perilla seed oil group at the high fat level. The stimulating responses of lymphycotes by PHA did not seem to be influenced by different dietary fat sources. However, conA mitogenic responses was significantly increased in perilla seed oil group. The lower level of perilla seed oil (2%, 15%) showed slightly higher responses of ConA, indicating that lower level of perilla seed oil might have stimulatory response on the immune response. The number of antibody forming cells of spleen against SRBC was increased in 30% fat level for all the three kind of fats. However, no effect has been found in plaque forming cell response by the differences in dietary fat sources. There were no significant differences in serum IgG and IgA levels in all dietary groups.

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Evaluation of Antibodies Against Haemophilus influenzae Type b in Korean Adults (우리나라 성인에서 Haemophilus influenzae type b에 대한 항체 평가)

  • Lee, Ji Hyen;Kim, Han Wool;Lee, Soyoung;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.24 no.3
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    • pp.125-133
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    • 2017
  • Purpose: After the introduction of Haemophilus influenzae type b (Hib) vaccine in 1995 in Korea, it was included in the national immunization program in 2013. In the post-Hib vaccine era, some studies in other countries reported that invasive Hib disease affects adults, especially the elderly and immunocompromised persons, more often than it affects children. To evaluate disease susceptibility, quantitative and qualitative analysis of anti-polyribosylribitol phosphate (PRP) antibodies were carried out in Korean adults aged 20 to 85 years. Methods: Sera were collected from 39 healthy adults (20 to 50 years of age) and from 30 elderly adults (75 to 85 years of age) who did not have immune-compromising conditions. The concentration of anti-PRP immunoglobulin G (IgG) and serum bactericidal indices (SBIs) were measured by enzyme-linked immunosorbent assay and serum bactericidal assay. Results: Geometric mean concentrations of anti-PRP IgG and geometric mean SBIs were $0.88{\mu}g/mL$ (95% confidence interval [CI], 0.17 to 3.85) and 354 (95% CI, 50 to 2,499) in young adults and $1.67{\mu}g/mL$ (95% CI, 0.53 to 5.24) and 449 (95% CI, 146 to 1,376) in elderly adults, respectively. When the threshold of seropositivity for anti-PRP IgG was applied as 0.15 or $1.0{\mu}g/mL$, which is the protective antibody level in children, seropositive rates were 87.2% or 53.8% in young adults and 100% or 60% in elderly adults. The seropositivity rates of the SBI ($SBI{\geq}4$) were 82.1% and 100% in the groups, respectively. Conclusions: Most subjects in the adult and elderly adult groups display immunity to Hib based on quantitative and qualitative antibody levels, but not all. Because high immunization and low Hib circulation rates may reduce the natural Hib immunity in the population, monitoring Hib immunity as well as disease are needed continuously.

Effect of dietary supplementation of Sargassum meal on laying performance and egg quality of Leghorn layers

  • Fan, Geng-Jen;Shih, Bor-Ling;Lin, Hui-Chiu;Lee, Tzu Tai;Lee, Churng-Faung;Lin, Yih-Fwu
    • Animal Bioscience
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    • v.34 no.3_spc
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    • pp.449-456
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    • 2021
  • Objective: Seaweeds could be an alternative and functional feed resource. The purpose of this experiment is to investigate the effect of dietary supplementation of Sargassum meal on laying performance and egg quality of layers. Methods: Two hundred 36-wk-old layers were divided into five treatment groups. Each treatment had four replicates with 10 hens per experimental unit. The corn-soybean meal basal diet was formulated as control group. Sargassum meals were included 0%, 1%, 2%, 3%, or 5% to diets for five treatment groups, respectively. Treatment groups were isocaloric-isonitrogenous diets. Laying performance and egg quality were measured for eight weeks. Results: Sargassum meal supplementation did not affect daily feed intake. Supplementation 1% to 3% of Sargassum meal in diets increased daily laying rate and egg mass compared with those from control group (p<0.05). Egg qualities among five groups were all similar. Supplementation of 3% Sargassum meal increased the lightness of egg yolk (p<0.05). Eggs produced from layers fed 1% and 2% Sargassum meal had a higher consumer's acceptability than the control group (p<0.05). In blood characteristics, contents of glucose, nitrogen, triiodothyronine (T3) and thyroxine (T4) increased as the increase of supplementation ratio of Sargassum meal (p<0.05). In serum antibody titers, supplementation of 2% Sargassum meal stimulated a higher immunoglobulin M (IgM) level than that from control group (p<0.05). However, IgM content of layers fed diets with Sargassum meal ≥3% were decreased (p<0.05). There was no difference in IgA and IgG titers among groups. Conclusion: Supplementation of 1% to 3% Sargassum meal has shown to increase egg laying rate and egg mass of Leghorn layers. However, high supplementation (5%) would negatively affect laying performance. In consideration of laying performance, egg quality, consumer responses, and blood antibody, supplementation of Sargassum meal was suggested 2% in the diet for layers.

Effects of Gal-13 on the Content of Immunoglobulin, Proliferation of Lymphocyte and Antibody Titers after Vaccination with Infectious Bursal Disease Virus Vaccine in Chickens

  • Yang, Yurong;Jiang, Yibao;She, Ruiping;Peng, Kaisong;Zhou, Xuemei;Yin, Qingqiang;Wang, Decheng;Liu, Tianlong;Bao, Huihui
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.3
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    • pp.405-411
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    • 2007
  • Gal-13 is an antimicrobial peptide isolated from chicken intestine. Ninety chickens were randomly divided into two groups (45 chickens for each group) to determine the effect of oral administration of Gal-13 on the acquired immune response. The chickens in the first group were fed a diet without Gal-13 as the control, and the chickens in the second group were fed the same diet, except that Gal-13 ($1{\mu}g/ml$) was suspended in drinking water just after hatching. Samples of blood, thymus, bursa of fabricius and spleen were taken at day 1, 4, 7, 10 and 17. The chickens in both groups received infectious bursal disease virus vaccine at day 20, and then sera samples were collected for analysis at 14, 21, 28 and 35 days after vaccination. The results showed: (1) Gal-13 could enhance the content of immunoglobulin (Ig)G at the age of 4 to10 days (p<0.05) and IgM at the age of 4 and 10 days (p<0.05) in the serum; (2) In vitro experiments showed that Gal-13 (0.625-1.250${\mu}g/ml$) enhanced the proliferation of peripheral blood lymphocytes of the chickens stimulated by lipopolysaccharide (LPS) and concanavlin A (ConA). Compared to the control, Gal-13 (1 ${\mu}g/ml$) enhanced the proliferation of bursa lymphocytes at 17 days of age (p<0.01) and thymus lymphocytes at 7 days of age (p<0.01), but restrained lymphocyte proliferation in chicken spleen and differed significantly at day 10 (p<0.01); (3) Gal-13 enhanced infectious bursal disease virus antibody in sera of chickens 21 days after infectious bursal disease virus vaccine administration (p<0.05). These results suggested that Gal-13 could modulate adaptive immune responses of chickens.