• 한국미생물·생명공학회지
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• 제16권5호
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• pp.335-342
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• 1988

하이브리도마를 쥐의 복강이나 in-vitro에서 배양한 뒤 생산된 IgG1 type의 쥐 단일클론 항체를 정제하기 위하여 음이온 교환 크로마토그래피를 이용하였다. 배양이 끝난 배지를 원심분리하여 세포를 제거하고 50-60% ammonium sulfate로 침전물을 만든 다음 0.025M Tris-HCI(pH8.2)용액으로 투석하여 salt가 제거된 sample을 DEAE-Trisacryl M에 부하하였다. Column에 결합된 항체는 30-40mM NaCl 을 포함하는 0.025M Tris-HCI(pH8.2)용액으로 용출하였다. 혈청농도가 높은 배지 (10% FBS)에서는 50% ammonium sulfate 처리로 90% 이상의 항체가 회수되었으나 저혈청 배지 (2% FBS)에서는 60% ammonium sulfate 처리에도 회수율이 84%에 그쳤다. 후자의 경우 한외여과법 (ultrafiltration)을 이용하여 항체 회수율을 91%까지 증가시킬 수 있으나 농축된 항체를 크로마토그래피로 정제하였을 때 그순도가 ammonium sulfate 침전법에 비해 낮아졌다. 하이브리도마 Alps 25-3, HCGK, A4W, KW를 여러가지 배양조건에서 배양한 뒤 생산된 항체를 DEAE-Trisacryl M chromatography를 이용하여 정제해 본 결과 대체로 순도 70-80%의 항체를 얻을 수 있었고 이때 항체 회수율은 65% 선이었다. 항체의 순도를 높이기 위해서는 affinity chromatography 혹은 gel filtration과 같은 2차적인 방법이 필요 할 것으로 보이며 한 예로 affinity chromatography를 이용하여 순도 95%의 항체를 얻었다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권5호
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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.

• 대한수의학회지
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• 제9권2호
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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.

• 약학회지
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• 제39권5호
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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.

• 대한소아치과학회지
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• 제30권1호
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• pp.61-68
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• 2003

치아우식증을 예방하기 위한 백신연구를 위하여 주원인균인 Sterptococcus mutans 균체항원과 이 균에서 분리한 glucosyltransferase (GTF)를 항원으로 하고 이에 면역조절기능이 있는 retinoic acid (RA)를 첨가하여 투여경로와 백신조성이 이들 항원에 대한 면역반응에 미치는 영향을 실험하였다. 균체항원(Ingbritt strain)을 마우스의 피하에 Complete Freund's Adjuvant와 함께 투여하여 생산되는 혈청내 응집항체가는 serotype e (LM-7)와는 강한 교차반응을 보였으나 serotype f (OMZ-175)와는 거의 교차반응을 일으키지 않았다. 면역혈청내 항-GTF 및 항-Ag I/II 항체중 항-GTF IgA는 피하로 투여시 전혀 검출되지 않았으나 이에 RA를 첨가하면 다량의 항체 생산을 관찰하였고 그 정도는 경구투여시의 생산량을 능가하였다. GTF를 alum과 함께 투여하여 생산되는 혈청내 항-GTF 항체중 IgM은 피하로 투여시 상당량이 검출되었고 RA를 첨가하면 그 생산이 증가되었으며 경구로 투여시 대조군에 비하여 약간증가를 보였으나 피하투여시의 그것에는 미치지 못하였다. GTF-특이 IgG는 경구투여시는 전혀 검출되지 아니하였고, 피하투여시에만 현저한 증가를 보였으며, RA첨가는 이에 영향을 미치지 못하였다. 항-GTF IgA는 피하로 투여시 전혀 검출되지 아니하였으나 이에 RA를 첨가하면 증가된 항체생산을 관찰하였고 그 정도는 경구투여시의 생산량을 능가하였다. 이상의 실험성적은 GTF에 대한 항체생산은 투여경로와 항원의 종류에 따라 다양한 반응을 나타내며 RA는 이를 백신에 첨가하면 피하경로를 이용하여 면역하더라도 경구투여와 유사한 IgA-매개 면역반응으로 조절시킬 수 있는 가능성을 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제24권5호
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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.

• Journal of Nutrition and Health
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• 제20권5호
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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.

• Pediatric Infection and Vaccine
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• 제24권3호
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• pp.125-133
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• 2017

목적: 1995년 한국에서 b형 헤모필루스 인플루엔자(Haemophilus influenzae type b [Hib]) 백신이 도입된 이후, 2013년 국가예방접종에 포함되었다. Hib 백신 도입 시기 이후 다른 나라의 일부 연구에 따르면, 침습적 Hib 질환은 소아보다 성인, 특히 노인 및 면역 저하 환자에서 더 자주 발생한다고 보고하였다. 따라서, 본 연구는 20-85세의 한국 성인에서 항-polyribosylribitol phosphate (PRP) 항체의 양적 및 질적 측정을 통하여 질병 감수성을 평가하고자 하였다. 방법: 혈청은 39명의 건강한 성인(20-50세)과 면역 저하 상태가 아닌 30명의 노인(75-85세)에서 수집하였다. 모든 피험자는 7일 이내에 항생제를 복용하지 않았다. 항-PRP 면역글로불린 G (immunoglobulin G [IgG]) 항체가 및 혈청 살상능 역가(serum bactericidal index [SBI])는 효소결합면역흡착측정법과 혈청 살상능 평가분석을 통해 측정하였다. 결과: 항-PRP IgG 항체가의 기하평균 및 SBI의 기하평균은 성인군에서 각각 $0.88{\mu}g/mL$ (95% 신뢰 구간, 0.17-3.85) 및 354 (95% 신뢰구간, 50-2,499)이었고, 노인군에서는 각각 $1.67{\mu}g/mL$ (95% 신뢰구간, 0.53-5.24) 및 449 (95% 신뢰구간, 146-1,376)이었다. 소아에서 Hib 감염에 대한 방어 농도로 인정하고 있는 0.15 및 $1.0{\mu}g/mL$을 적용하였을 때, 항체 양성률은 성인군에서 87.2%, 노인군에서 100%였다. SBI의 양성률($SBI{\geq}4$)은 성인군에서 82.1%, 노인군에서 100%였다. 결론: 대부분의 성인군과 노인군에서 양적으로나 기능적으로 Hib 질환에 대한 방어 면역력을 가지고 있었으나 모두는 아니었다. 높은 면역력과 낮은 Hib 순환율은 인구에서 Hib에 대한 자연면역을 감소시킬 수 있으므로, Hib 면역과 질병에 대한 지속적인 감시가 필요하다.

• Animal Bioscience
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• 제34권3_spc호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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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.