• Title/Summary/Keyword: ELISA antibody

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Hapten Synthesis and Influence of Coating Ligands on Enzyme-linked Immunoreaction of DDT

  • Hong, Ji- Youn;Kim, Jong-Hyun;Choi, Myung-Ja
    • Bulletin of the Korean Chemical Society
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    • v.23 no.10
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    • pp.1413-1431
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    • 2002
  • For the development of immunodetection method of 4,4'-dichlorodipheny-2,2,2-trichloroethane (p,p'-DDT), a persistent and broad toxic organochlorine insecticide, various DDT derivatives were synthesized and characterized for the use of immunogens and the coating ligands for the antibody evaluation. The appropriate lengths of linkers were introduced to investigate more efficient DDT derivatives. Among these hapten derivatives, 2,2-Bis(4-chlorophenyl)acetic acid (DDA), 5,5-Bis(4-chlorophenyl)-5-hydroxypentanoic acid (DDHP) and 5,5-Bis(4-chlorophenyl)-5-chloropentanoic acid (DDCP) were conjugated with keyhole limpet hemocyanin (KLH) for the use of immunogen to produce antibodies. 6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoic acid (DDHH) and 3-[6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoylamino]propanoic acid (DDHHAP) in addition to above hapten derivatives were conjugated to ovualbumin (OVA) and bovine serum albumin (BSA) for the use of coating ligands to measure the titration level of antibody and the displacement of free analytes. Three matching pairs of antibodies and coating ligands were selected for the simultaneous detection of p,p'-DDT and its related compounds of DDA and 2,2-bis(4-chlorophenyl)-1,1-dichloroethylene (p,p'-DDE) by investingating the displacement of free analytes in an indirect ELISA. These were PAb #1 and coating ligand DDCP-OVA, PAb #1 and DDHHAP-OVA, and PAb #3 and DDHHAP-OVA. The most useful immunoreaction for DDT analytes were obtained using PAb #3 and coating ligand DDHHAP-OVA showing 3.4 ng/mL of lower limit of detection. These results indicated that titration level and free analytes displacement were greatly influenced by hapten derivatized and carrier proteins conjugated.

Different immunological features of two genetically distinct type 2 porcine reproductive and respiratory syndrome (PRRS) viruses

  • Shabir, Nadeem;Khatun, Amina;Kim, Won-Il
    • Korean Journal of Veterinary Service
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    • v.37 no.1
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    • pp.1-9
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    • 2014
  • Although it has been generally accepted that porcine reproductive and respiratory syndrome virus (PRRSV) induces weak and delayed protective immunity after infection, it is unclear that the same immunological features can be applicable to all PRRS viruses because huge genetic variation exists even among the same genotypes of PRRSV (Type 1 and 2). In the current study, two genetically distinct type 2 PRRSV strains (VR-2332 and JA142) which showed approximately 90% nucleotide homology based on ORF5 sequences were characterized by both in vitro and in vivo assessments to determine the immunological features of the viruses. For in vitro assessment, porcine alveolar macrophages (PAM) were infected with the viruses at $10^{-3}$ multiplicity of infection (MOI) and then supernatants and cells were collected separately at 36 hrs post infection to determine the relative expression levels of IL-$1{\alpha}$, IL-12, TNF-${\alpha}$ and INF-${\alpha}/{\beta}$ by quantitative RT-PCR. In addition, five PRRSV-free pigs were inoculated with either of JA142 or VR2332 for in vivo assessment. Serum samples were collected every week until 6 weeks post challenge. The serum samples were analyzed for the levels of viremia, PRRSV nucleocapsid-specific antibody and virus neutralizing antibody. Based on those assessments, the two viruses showed different patterns of cytokine expression in PAM and immune responses in pigs after infection. These results indicate that genetically distinct PRRSV strains have different immunological features, which might be criteria for virus classification and selection of candidate virus strains for vaccine development in the future.

Production of Monoclonal Antibody Against Listeria monocytogenes and Its Application to Immunochromatography Strip Test

  • Shim, Won-Bo;Choi, Jin-Gil;Kim, Ji-Young;Yang, Zheng-You;Lee, Kyu-Ho;Kim, Min-Gon;Ha, Sang-Do;Kim, Keun-Sung;Kim, Kwang-Yup;Kim, Cheol-Ho;Ha, Kwang-Soo;Eremin, Sergei A.;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.17 no.7
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    • pp.1152-1161
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    • 2007
  • An immunochromatography (ICG) strip test based on a monoclonal antibody for the rapid detection of L. monocytogenes in meat and processed-meat samples was developed in this study. A monoclonal antibody (MAb) specific to L. monocytogenes was produced from cloned hybridoma cells (FKLM-3B12-37) and used to develop an ICG strip test. The antibody showed a stronger binding to L. monocytogenes than other Listeria species, and a weak cross-reaction to S. aureus based on an ELISA. The detection limit of the ICG strip test was $10^5\;cell/ml$. In total, 116 meat and processed-meat samples were collected and analyzed using both the ICG strip test and a PCR. The ICG strip test and PCR indicated L. monocytogenes contamination in 34 and 27 meat samples, respectively. The 7 meat samples not identified as L. monocytogenes positive by the PCR were also tested using an API kit and found to be contaminated by Listeria species. In conclusion, the ICG strip test results agreed well with those obtained using the PCR and API kit. Thus, the developed ICG has potential use as a primary screening tool for L. monocytogenes in various foods and agricultural products, generating results within 20 min without complicated steps.

Sparganum infections in normal adult population and epileptic patients in Korea: A seroepidemiologic observation (항체검사에 의한 한국인 스파르가눔 감염의 혈청역학적 조사)

  • Yoon Kong;Seung-Yull Cho;Woo Shik Kang
    • Parasites, Hosts and Diseases
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    • v.32 no.2
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    • pp.85-92
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    • 1994
  • A seroepidemiologic observation of anti-Spirometrc erinacei plerocercoid (sparganum) antibody (IgG) in serum was made in normal adult and epileptic patients in Korea from february, 1987 to September, 1990. Sera were tested by enzyme-linked immunosorbent assay (ELISA) for anti-sparganum antibody together with anti-Taenic soEiun metacestode, and anti-Parusonimus westermcni antibodies. Sera reacted positively to sparganum antigen only were considered. Positive rate for anti-sparganum antibody in 850 normal adults was 1.9% (standardized rate by provincial population was 1.7%). In 2,667 randomly selected patients of epilepsy at 28 local centers of the Changmi Club, positive rate was 2.5% (standardized rate: 2.3%). In both normal adult and patient groups, the higher antibody rates were observed in Kangwon and Chonnam Provinces. Positive rates were 10 times higher in male than in female in normal adults and 4.5 times in male epileptic patients. The rates were elevated especially with age over 30-year. odd ratio of the antibody was 1.32 which indicated an ambiguous etiologic factor for epilepsy.

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Immune Reaction of the Vaccinated Hamsters with Combined Hantaan-Puumala Vaccine (신증후출혈열의 혼합백신을 접종한 햄스터에서의 면역성 조사)

  • Lee, Ho-Wang;Chu, Yong-Kyu;Cui, Long-Zhu;Woo, Young-Dae;Ahn, Chang-Nam;Kim, Hoon;Jang, Yang-Seok
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.39-47
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    • 1997
  • A large number of viruses belonging to Genus Hantavirus in Family Bunyaviridae are etiologic agents for hemorrhagic fever with renal syndrome (HFRS), or hantavirus pulmonary syndrome (HPS). Hantaan (HTN), Seoul (SED), Belgrade (BEL), Puumala (PUU) serotype viruses are well known causative agents for HFRS in Eurasian continent. Among those viruses Hantaan and Seoul serotypes are well known to cause HFRS in Korea, but there are some sporadic incidence by other than Hantaan or Seoul viruses. Recently we have developed the combined Hantaan-Puumala virus vaccine to prevent world-wide occuring HFRS. This combined vaccine is formalin inactivated, suckling mouse and suckling hamster brain extracts for Hantaan and Puumala viruses, respectively. Protein contents of this purified candidate vaccine is $27\;{\mu}g/ml$, which contains 1,024 ELISA antigen units to each virus, but content of myelin basic protein which is causing experimental allergic encephalomyelitis is less than 0.1 ng/ml. Thirty hamsters were given twice at one month interval intra-muscularly and bled on 30 days after each vaccination from retro-orbital sinus vein. Antibody titers were tested against 5 major serotype viruses, Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses by IFA and PRNT. The mean IF antibody titers on 30 days after primary shot were 78.4, 68.8, 68.8, 37.9, and 15.6; mean neutralizing antibody titers were 65.4, 12, 6.1, 65.6 and 0.5 against Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses, respectively. The mean IF antibody titers on 30 days after booster shot were 686.9, 567.5, 550.4, 516.3, and 430.9; and neutralizing antibody titers were 710.8, 41.9, 24.3, 409.9, and 1.6 against Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses, respectively.

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A Monoclonal Anti-peptide Antibody against $\beta$2-adrenergic Receptor Which Specifically Binds [$^{3}H$] dihydroalprenolol

  • Shin, Chan Young;Noh, Min Su;Lee, Sang Derk;Lee, Sang Bong;Ko, Kwang Ho
    • Biomolecules & Therapeutics
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    • v.3 no.4
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    • pp.266-272
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    • 1995
  • The analysis of membrane receptors for hormones and neurotransmitters has progressed considerably by pharmacological and biochemical means and more recently through the use of specific antibodies. To generate and characterize a moloclonal antibody against $\beta$-adrenergic receptor, a synthetic $\beta$2-adrenergic receptor peptide (Phe-Gly-Asn-Phe-Trp-Cys-Phe-Trp-Thr-Ser-lle-Asp-Val-Leu) which may comprise part of $\beta$-adrenergic receptor ligand binding pocket was coupled to Keyhole Limpet Hemocyanin (KLH) and used as an immunogen. Male BALB/C mice were immunized with this antigen and the immunized spleen was fused with myeloma SP2/0-Ag14 cells to produce monoclonal antibodies. Two clones were obtained but one of monoclonal antibodies, mAb5G09, was used throughout in this study because the other clone, mAb5All showed weak immunoreactivity against KLH as well. The mouse monoclonal antibody mAb5G09 produced in this study showed immunoreactivity to peptide-KLH conjugates and also to human A43l cells and guinea pig lung $\beta$2-adrenergic receptor as revealed by ELISA and western blot. In the course of determination of the effects of mAb5G09 on $\beta$-receptor ligand binding, it was observed that mAb5G09 specifically bound $\beta$-adrenergic radioligand [$^3$H]dihydroalprenolol (DHA) with a dissociation constant (Kd) of 60 nM. The [$^3$H]DHA binding activity of mAb5G09 had characteristics of immunoglobulins and the binding activity was not observed in the control anti-KLH monoclonal antibody. The monoclonal antibody, mAb5G09 produced in this study may provide useful models for the study of the structure of receptor binding sites.

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A sere-epidemiological survey on Ibaraki disease in western area of Kyunggi province (경기 서부지방의 소 Ibaraki병 중화항체가 조사)

  • 이우종;고신일;최영래;강영배;최강석
    • Korean Journal of Veterinary Service
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    • v.19 no.3
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    • pp.207-211
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    • 1996
  • To investigate the serum neutralizing antibodies against Ibaraki disease virus in the western area of Kyunggi province, a sero-epidemiological survey was done from August 1995 to March 1996. The results obtained are summarized as follows : 1. An overall prevalence of the neutralizing antibodies agaist Ibaraki virus was as high as 68.6% (218 positive reactors amomg 318 heads of dairy cattle). 2. It showed the regional differences with 60.5${\ulcorner}$(46/76) in Koyang, 75.2% (100/133) in Paju and 66.1% (72/109) in Kimpo. 3. It also appeared with a seasonal difference showing 74.4% of prevalence with the mean titer higher than 60 during the mosquito season (from August to November) and 58.6% of prevalence with the mean titer 22 after the mosquito season to March. 4. Any cross reactions between Ibaraki and bluetongue viruses were not detected in the ELISA and AGID tests.

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Detection of Paragonimus-specific IgG antibody in CSF and pleural effusion by micro-ELISA (면역효소진단법에 의한 뇌척수액 및 흉막삼출액에서의 폐흡충 특이 IgG항체 검출)

  • 조승열;김성일
    • Parasites, Hosts and Diseases
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    • v.21 no.2
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    • pp.286-288
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    • 1983
  • 폐흡충 감염자에서 나타나는 중추신경계의 병변을 모두 폐흡충이 중추신경계를 침범하여 발생한 것이라고는 할 수 없다. 그러므로 이 경우 원인진단을 위해서는 뇌척수액에 나타나는 폐흡충 특이항체의 측정이 필요하다고 생각된다. 우리는 확인된 뇌폐흡충중 2례와 척수 스파르가눔증 1례, 뇌병변이 없는 폐흡충증 환자 1례와, 기타 중추신경계 질환 환자 10례에서 얻은 뇌척수액을 희석하지 않고 면역효소진단법으로 특이항체를 측정하였다. 그 결과 흡광도 0.25를 양성 기준으로 하면 뇌 폐흡퉁증을 진단할 수 있다고 생각하게 되었다. 폐흡충중 환자 2례의 흉막삼출액에서 특이 IgG 항체가는 혈청에서의 측정치와 다르지 않아, 흉막삼출액도 폐흡충증의 진단에 이용할 수 있다고 생각되었다.

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Immune reactions and allergy in experimental anisakiasis

  • Cho, Sung-Weon;Lee, Haneul-Nari
    • Parasites, Hosts and Diseases
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    • v.44 no.4 s.140
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    • pp.271-283
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    • 2006
  • The third-stage larvae (L3) of the parasitic nematode, Anisakis simplex, have been implicated in the induction of hyperimmune allergic reactions in orally infected humans. In this work, we have conducted a review of an investigation into immune reactions occurring in animals experimentally infected with A. simplex L3. The patterns of serum antibody productions if the experimental animals against excretory-secretory products (ESP) of A. simplex L3 contributed to our current knowledge regarding specific humoral immune reactions in humans. In our review, we were able to determine that L3 infection of experimental animals may constitute a good model system for further exploration of immune mechanisms and allergy in anisakiasis of humans.

Enhancement of nerve growth factor production and release by buthanol fraction of Liriope platyphylla in C6 cells and rat cultured astrocyte

  • Hur, Jin-Young;Lee, Pyeong-Jae;Kim, Jeong-Min;Kim, Ho-Cheol;Kim, Sun-Yeou
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.135.3-136
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    • 2003
  • Liriope platyphylla (LP) Wang et Tang has been used for tonic, anti-tussive and expectorant in Korea. In the current study, we found that buthanol fraction of Liriope platyphylla-conditioned media of C6 and primary astrocyte induced the neurite outgrowth of PC 12 cells, which effect was reversed by addition of NGF-antibody. We demonstrated that buthanol fraction of Liriope platyphylla increased the expression and secretion of NGF through RT-PCR and ELISA. (omitted)

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