• Title/Summary/Keyword: ELISA IgM

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Changes in blastogenic responses and antibody titers of mice infected with Toxoplasma gondii (Toxoplasma gondii 감염 마우스에 있어서의 아세포화 반응 및 항체가 변화)

  • 신대환;이영하
    • Parasites, Hosts and Diseases
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    • v.30 no.2
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    • pp.125-132
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    • 1992
  • This study was performed to observe the cell-mediated and humoral immune responses in mice which were infected with Beverley, Fukaya and ME49 strain of Toxoplasma gondii, respectively. The blastogenic responses of splenocytes using $[^3H]-thymidine$ and serum antibody titers were measured weekly up to 10 weeks after infection. The blastogenic responses of splenocytes treated with concanavalin A and Tosoplasma Iysate were significantly declined in the 3 strain groups as compared with the non-infected group (P<0.05), however lipopolysaccharide-treated blastogenic responses were not significantly different between infected and non.infected groups. The serum IgG antibody titers in the three infected groups increased from 2 weeks after infection, and the serum IgM antibody titers increased until 4 weeks after infection. No significant differences were revealed in blastogenic responses and serum antibody titers among the 3 groups. The present study suggested that cell-mediated immune responses were involved in T. gondii infected mice and blastogenic responses of T Iymphocytes were inhibited in acute T. gondii infection.

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A Study on the Effect of Conjugated Linoleic Acid on Immunosuppressed Mice with Corticoids

  • 임형진;김진영;임좌진;이병한;박희명;정병현
    • Journal of Veterinary Clinics
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    • v.20 no.1
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    • pp.59-65
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    • 2003
  • This study was performed to investigate the immunopotentiative effects of conjugated linoleic acid on mice immunosuppressed by administratin corticoids. Mice were divided into four groups of 8 mice. Two groups (C, CP) were given diet supplemented with 1% linoleic acid (CLA) and the other two groups (L, LP) were given diet supplemented with 1% linoleic acid (LA) instead of CLA. Prednison was administered to two groups (CP, LP) for immune depression. After feeding diets for 3 weeks containing PDS injection for last 1 week. Serum and gut lumen lavage were taken. Measurement of total Ig were executed using sandwich ELISA. Serum levels of IgA, IgG, and IgM showed some trend which groups fed with CLA were higher than groups fed with LA while IgE was reduced in those fed the CLA intake, and groups administered with PDS were lower than groups administered with saline. However, no significant differences were seen in the proportion of total immunoglobulin in serum. In case of secretory IgA, Group C and CP were significantly higher than group L and LP. Especially between CP and LP, it can be seen effects of CLA. In addition that the CLA treated group weighted a significantly lower level than the one's that have not been treated with CLA. These result support the view that CLA potentiate the immune response and prevent immune depression caused by administrating of corticoids. In conclusion, CLA produced a situation favorable for immunopotentiative effects. Thus, the clinical application of CLA is warranted.

Lipopolysaccharide Yields from Rhodobacter capasulatus with indirect ELISA

  • Yoo, Tae-Eun;Lee, Hyun-Soon
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.255-262
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    • 1996
  • The lipopolysaccharide (LPS) yields were measured in Rhodobacter capsulatus under several conditions by the ELISA method. The purification of LPS was done by affinity chromatography of IgG coupled CNBr-activated sepharose-4B instead of ultra-centrifugation. The purity of the LPS didn't show much difference between affinity chromatography and ultra-centrifugation method, but affinity chromatography method required much fewer organisms and was more convenient. LPS yield was measured in ng units by the ELISA method. Mannitol was a better single carbon source than other sugars, but mixing two carbon sources resulted in greater LPS yields than any sugar alone. LPS yield was directly proportional to $NH_ 4CI$ concentration, with optimum yields at 0.05% nitrogen. In contrest to LPS yields, which decreased at 0.005% nitrogen concentration total protein was increased 16 times. Calcium influenced LPS yields. At 0.7 mM $CaCI_ 2$, the LPS yield was 16.5 $\mu$g/mg DW, five times the yield without calcium.

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Production of Group Specific Monoclonal Antibody to Aflatoxins and its Application to Enzyme-linked Immunosorbent Assay

  • Kim, Sung-Hee;Cha, Sang-Ho;Karyn, Bischoff;Park, Sung-Won;Son, Seong-Wan;Kang, Hwan-Goo
    • Toxicological Research
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    • v.27 no.2
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    • pp.125-131
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    • 2011
  • Through the present study, we produced a monoclonal antibody against aflatoxin B1 (AFB1) using AFB1-carboxymethoxylamine BSA conjugates. One clone showing high binding ability was selected and it was applied to develop a direct competitive ELISA system. The epitope densities of AFB1-CMO against BSA and KLH were about 1 : 6 and 1 : 545, respectively. The monoclonal antibody (mAb) from cloned hybridoma cell was the IgG1 subclass with ${\lambda}$-type light chains. The $IC_{50}s$ of the monoclonal antibody developed for AFB1, AFB2, AFG1 and AFG2 were 4.36, 7.22, 6.61 and 29.41 ng/ml, respectively, based on the AFB1-KLH coated ELISA system and 15.28, 26.62, 32.75 and 56.67 ng/ml, respectively, based on the mAb coated ELISA. Cross-relativities of mAb to AFB1 for AFB2, AFG1 and AFG2 were 60.47, 65.97 and 14.83% in the AFB1-KLH coated ELISA, and 59.41, 46.66 and 26.97% in the mAb coated ELISA, respectively. Quantitative calculations for AFB1 from the AFB1-Ab ELISA and AFB1-Ag ELISA ranged from 0.25 to 25 ng/ml ($R^2$ > 0.99) and from 1 to 100 ng/ml ($R^2$ > 0.99), respectively. The intra- and inter-assay precision CVs were < 10% in both ELISA assay, representing good reproducibility of developed assay. Recoveries ranged from 79.18 to 91.27%, CVs ranged from 3.21 to 7.97% after spiking AFB1 at concentrations ranging from 5 to 50 ng/ml and following by extraction with 70% methanol solution in the Ab-coated ELISA. In conclusion, we produced a group specific mAb against aflatoxins and developed two direct competitive ELISAs for the detection of AFB1 in feeds based on a monoclonal antibody developed.

Production of monoclonal antibodies to immunoglobulin M of sevenband grouper (Epinephelus septemfasciatus) (능성어(Epinephelus septemfasciatus) immunoglobulin M에 대한 단클론 항체 생산)

  • Kim, Si-Woo;Kim, Jong-Oh;Kong, Kyoung-Hui;Oh, Myung-Joo;Kim, Wi-Sik
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.111-115
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    • 2021
  • Immunoglobulin M (IgM) of sevenband grouper (Epinephelus septemfasciatus) was purified by mannan-binding protein (MBP) affinity column. The purified IgM had an apparent molecular weights of 76 (heavy chain) and 28 (light chain) kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Eight hybridoma clones secreting monoclonal antibodies (mAbs) against sevenband grouper IgM were established. Antibody detection enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the 8 mAbs revealed that optical density (OD) values were clearly different between sera from BSA-immunization and non-immunization of sevenband grouper. These results suggest that the produced mAbs in this study are specifically reacted with IgM of sevenband grouper.

Cytomegalovirus Infection among Pregnant Women in Beijing: Seroepidemiological Survey and Intrauterine Transmissions

  • Jin, Qing'e;Su, Jianrong;Wu, Shanna
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.1005-1009
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    • 2017
  • Primary cytomegalovirus (CMV) infection during pregnancy can cause congenital defects. Available data for CMV infection during pregnancy in north China are inadequate. The aim of this study was to evaluate the epidemiology of maternal CMV infection and explore the incidence of congenital infection. In this prospective study, serum CMV IgG and IgM antibodies were measured in 2,887 pregnant women using ELISA, and the IgG avidity test was performed on all IgM-positive subjects. The seroprevalence of anti-CMV IgG was 94.70%, and of anti-CMV IgM was 1.28%. CMV IgG prevalence increased significantly with age (p < 0.01). Women living in downtown areas showed higher IgG prevalence than those residing in urban areas (p = 0.023). CMV-IgM seroprevalence was highest in autumn (p = 0.021). There was no difference in IgM seroprevalence by age, socioeconomic status, geographical area, or gravida. The rate of primary CMV infection was 0.45% (13/2,887) at the first trimester. The seroconversion rate during pregnancy was 0.76% (22/2,887). One woman underwent seroconversion during pregnancy and gave birth to an infant with asymptomatic CMV infection. Congenital CMV infection was diagnosed in five of the 14 infants from 14 mothers with active infection, for a vertical transmission rate of 35.71% (5/14). Three infants were asymptomatic, whereas two infants presented symptomatic infection with hearing deficits. Although CMV IgG prevalence is relatively high in north China, significant attention to primary CMV infection during pregnancy is still needed.

Production of Monoclonal Antibodies Specific to Korean Mistletoe pectin (KML-C) and Their Characterization (한국산 겨우살이 렉틴 (KML-C)에 대한 단일크론항체의 생산과 특성)

  • 윤택준;유영춘;강태봉;김성훈;김갑수
    • YAKHAK HOEJI
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    • v.45 no.2
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    • pp.180-189
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    • 2001
  • We have reported that water-extracted Korean mistletoe (KM-110) had various biological activities such as antitumor and immunomodulatory activity, and the pectin fraction (KML-C) of the extract was one of major factors related to its biological functions. In this paper, we produced murine monoclonal antibody (mAb) against KML-C. The cAbs obtained were largely classified into two groups according to specificity to KML-C and ML-I, a pectin from European mistletoe. One group mAbs (9H7-D10 and 3C2-lH4) strongly reacted with KML-C, but not ML-I. In contrast, another group cAbs (8Bll-2C5, BE12-3E9 and 5E10-Fl) reacted with both KML-C and ML-1. The subisotypes of these mobs were shown to be IgGl (9H7-lD10, 3C2-lH4 and 8Bll-2C5) or IgM (8E12-3E9 and 5E10-Fl). To develop an assay system for determination of the amount of KML-C, we established the sandwich ELISA (enzyme-linked immunosorbent assay) method using these mAbs and horse radish peroxidase (HRP)-labelled cAbs. In various combinations of the cAbs for coated antibody and detection antibody, the sandwich ELISA quantitatively detected KML-C, showing the detection limit ranging from 7-5,000 ng/ml. Especially reproducibility (C.V) of the sandwich ELISA, in which 8E12-3E9 was used for coating antibody and 8Bll-2C5-HRP for detection antibody, was 4.59-5.83 in intra assay, and 3.9-9.4 in inter assay.

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Changes of Binding Ability of Milk-Hypersensitive Patients질 IgE to Gamma-Irradiated Milk Proteins (감마선 조사된 우유단백질에 대한 우유 알러지 환자의 IgE 결합능의 변화)

  • 조경환;육홍선;이주운;이수영;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.3
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    • pp.505-509
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    • 2001
  • This study was carried out to evaluate the application of food irradiation technology as a method for reducing milk allergies. Bovine $\alpha$-casein, $\beta$-casein, $textsc{k}$-casein, $\alpha$-lactalbumin(ALA), $\beta$-lactoglobulin (BLG) and serum albumin (BSA) were used as model allergens of milk proteins and the proten solution (2.0 mg/mL) with 0.01 M phosphate buffered saline (pH 7.4) was irradiated at 3, 5 and 10 kGy. Using milk-hypersensitive patients IgE (MHP-IgE), the changes of binding ability to irradiated proteins were observed by competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA). Affinity of MHP-IgE to milk proteins was higher in ALA and BLG than that of other proteins. Standard curve to each non-irradiated protein could be made with MHP-IgE for quantifying milk allergens. Binding abilities of MHP-IgE to the irradiated proteins, however, decreased with different slopes of the standard curves. Sensitivity of gamma irradiation was higher in ALA and BLG than of other proteins. These results indicated that irradiation technology can be used to reduce the milk hypersensitivity.

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Development of a Rapid Diagnostic Test Kit to Detect IgG/IgM Antibody against Zika Virus Using Monoclonal Antibodies to the Envelope and Non-structural Protein 1 of the Virus

  • Kim, Yeong Hoon;Lee, Jihoo;Kim, Young-Eun;Chong, Chom-Kyu;Pinchemel, Yanaihara;Reisdorfer, Francis;Coelho, Joyce Brito;Dias, Ronaldo Ferreira;Bae, Pan Kee;Gusmao, Zuinara Pereira Maia;Ahn, Hye-Jin;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.56 no.1
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    • pp.61-70
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    • 2018
  • We developed a Rapid Diagnostic Test (RDT) kit for detecting IgG/IgM antibodies against Zika virus (ZIKV) using monoclonal antibodies to the envelope (E) and non-structural protein 1 (NS1) of ZIKV. These proteins were produced using baculovirus expression vector with Sf9 cells. Monoclonal antibodies J2G7 to NS1 and J5E1 to E protein were selected and conjugated with colloidal gold to produce the Zika IgG/IgM RDT kit (Zika RDT). Comparisons with ELISA, plaque reduction neutralization test (PRNT), and PCR were done to investigate the analytical sensitivity of Zika RDT, which resulted in 100% identical results. Sensitivity and specificity of Zika RDT in a field test was determined using positive and negative samples from Brazil and Korea. The diagnostic accuracy of Zika RDT was fairly high; sensitivity and specificity for IgG was 99.0 and 99.3%, respectively, while for IgM it was 96.7 and 98.7%, respectively. Cross reaction with dengue virus was evaluated using anti-Dengue Mixed Titer Performance Panel (PVD201), in which the Zika RDT showed cross-reactions with DENV in 16.7% and 5.6% in IgG and IgM, respectively. Cross reactions were not observed with West Nile, yellow fever, and hepatitis C virus infected sera. Zika RDT kit is very simple to use, rapid to assay, and very sensitive, and highly specific. Therefore, it would serve as a choice of method for point-of-care diagnosis and large scale surveys of ZIKV infection under clinical or field conditions worldwide in endemic areas.

Effects of Heat Treatment on the Antigenicity of Gamma-Irradiated Egg White Albumin (감마선 조사된 난백 알부민의 항원성에 미치는 가열의 영향)

  • 이주운;육홍선;조경환;김미리;김천제
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.848-853
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    • 2001
  • This research was conducted to study the effects on antigenicities (allergenicity) and structural changes of gamma-irradiated hen’s egg albumin (ovalbumin, OVA) by heating. Three groups of OVA solution (2.0 mg/mL) were prepared; 1) heat treatment; 2) irradiation after heating; 3) heating after irradiation. Samples were isothermally heated and/or irradiated at the absorption dose of 10 kGy. Competitive indirect ELISA was individually formatted with egg-allergic patients IgE (P-IgE), and mouse murine monoclonal IgG (M-IgG) and rabbit polyclonal IgG (R-IgG) for evaluating bindinhg abilities of antibodies to OVA in the sample solutions. Binding abilities of antibodies to thermally denatured OVA were changed : R-IgG to the sample treated with above 6$0^{\circ}C$, M-IgG to that above 7$0^{\circ}C$, and P-IgE to that above 8$0^{\circ}C$, respectively. P-IgE did not well recognize OVA heated at 8$0^{\circ}C$ and the above. However, binding abilities of M-IgG and R-IgG highly in creased. Significant differences of binding abilities were not observed in all samples with the combination of heat treatment and irradiation, regardless the order of the treatment. Turbidity of samples in creased both by heating and by irradiation, and the increase by irradiation was much higher than by heating. These results showed that allergenicity of OVA reduced by gamma irradiation was not affected by heating.

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