• Title/Summary/Keyword: ELISA antibody

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Serological survey of avian pneumovirus infection in laying hens of Gyeongbuk province (경북지역 산란계에서 avian pneumovirus 에 대한 항체가조사)

  • 김순태;김성국;조민희;김영환
    • Korean Journal of Veterinary Service
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    • v.26 no.1
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    • pp.51-56
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    • 2003
  • Avian pneumoviros(APV), also known as avian rhinotracheitis virus(ARTV), affects both turkeys and chickens and is known to be the primary causative agent of turkey rhinotracheitis (TRT). The aim of this study was to establish the presence or absence of antibodies to avian pneumovirus in the commercial poultry population of Korea. For this purpose, chicken serum samples were obtained and tested by an enzyme-linked immunosorbent assay (ELISA). The tested serum was collected in laying hens with reduction of egg production or normal in Gyeongbuk province. A total of 184 sera representing 42 different poultry farms of the Gyeongbuk region of Korea were included in this study. Laying hens of 16 different farms with reduction of egg production and laying hens of 26 different farms with clinically healthy at the time of serum sampling were considered positive to antibody against APV. In the farms with reduction of egg production, positive farm to antibody against avian pneumovirus were 11 of 16 different farms(68.8%) and positive sera were 47(58.8%) of 80 different serum. In the farms with clinically healthy flock, positive farm to antibody against avian pneumovirus were 12(46.2%) of 26 different farms and positive serum sample were 39(37.5%) of 104 different sera. According to the results tested to 42 different farms in 14 city, 8 of 14 city have flocks with antibody positive laying hens against APV, 1 of 14 city have antibody suspicious and 5 of 14 city shown antibody negative, respectively.

Development of an Enzyme-Linked Immunosorbent Assay Method for Residual Host Cell derived Proteins in Recombinant Antibody Drug Production (재조합 항체의약품의 생산시 생산세포주 유래 단백질 검출을 위한 ELISA 방법 개발)

  • Joung, Chan-Hi;Lim, Sang-Min;Koo, Yoon-Mo;Lee, Yong-Yoon;Son, Young-Su;Kim, Hyun-Il;Park, Heung-Rok
    • KSBB Journal
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    • v.21 no.3
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    • pp.212-219
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    • 2006
  • The purpose of this study was to develop a assay system of host cell-derived residual proteins in final pharmaceutical products. Accurate and simple assay system for host cell-derived proteins(HCPs) is very important test item in pharmaceutical qualification control. In this study, methods for quantification of residual HCPs in recombinant anti-GPIIbIIIa antibody were developed using a process-specific immunoligand assay which was based on the Enzyme linked Immunosorbent assay(ELISA) system. The assay had a detection limit of 10.8 ng/ml of HCPs with a product concentration of 1 mg/ml. The practical implication of these results is that the developed ELISA system can be used for HCPs qualification control and this system will be applicable to develop another ELISA system of different antibody drug.

Study on Antibody-enzyme Coupling and Enzyme Immunoassay Methods (효소-항체의 결합 및 효소면역측정 방법의 연구)

  • Jang Sean Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.3
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    • pp.874-879
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    • 2004
  • Alakaline phosphatase (ALP)- or horseradish peroxidase (HRP)-antibody conjugate was used frequently on the immunological detection methods such as enzyme-linked immunosobent assay (ELISA), immunobolt, immunohistochemistry. The classical enzyme-antibody coupling method by one-step (direction) injection of glutaraldehyde bring into being disadvantage such as low sensitivity of antigen detection because of homopolymers. This study was modified with the dialysis glutaraldehyde method to provide simple coupling through E-amino residues present in most protein. The dialysis glutaraldehyde coupling effects were better than the classical one-step glutaraldehyde injection in antigen detection of ELISA and immunobolt. Optimal dose of the dialysis glutaraldehyde solution was 0.10-0.25 %. This results suggest that the dialysis glutaraldehyde coupling method can readily applied to antigen detection of in vitro and in vivo.

Use of Serological-Based Assay for the Detection of Pepper yellow leaf curl Indonesia virus

  • Hidayat, Sri Hendrastuti;Haryadi, Dedek;Nurhayati, Endang
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.328-332
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    • 2009
  • Diseases caused by Pepper yellow leaf curl virus infection is considered to be emerging plant diseases in Indonesia in the last five years. One key factor for disease management is the availability of accurate detection of the virus in plants. Polyclonal antibody for Pepper yellow leaf curl Indonesia virus-Bogor (PYLCIV-Bgr) was produced for detection of the virus using I-ELISA and DIBA methods. The antibody was able to detect PYLCIV-Bgr from infected plants up to dilution 1/16,384 and cross reaction was not observed with Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV), and Chilli veinal mottle virus (ChiVMV). Positive reaction was readily detected in membrane containing Begomovirus samples from Yogyakarta (Kaliurang and Kulonprogo) and West Java (Bogor and Segunung). Infection of PYLCIV-Bgr in chillipepper, tomato, and Ageratum conyzoides was also confirmed using polyclonal antibody for PYLCIV-Bgr in DIBA. Polyclonal antibody for PYLCIV-Bgr is suggested to be included in disease management approach due to its good detection level.

Indirect ELISA Method for Measurement of Lactoperoxidase using IgY Antibody (IgY 항체를 이용하여 Lactoperoxidase 정량을 측정하기 위한 Indirect ELISA 방법의 개발)

  • 이승배;최석호;최재원
    • Food Science of Animal Resources
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    • v.24 no.2
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    • pp.182-188
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    • 2004
  • To determine the concentration of Lactoperoxidase (LPO), an indirect enzyme-linked immunosorbant assay(ELISA) was developed. Anti-LPO egg yolk immunoglobulin(IgY) was transferred to egg yolk by immunizing of Brown hens with LPO. The titer of purified anti-LPO IgY was 1: 520,000. The immunological response of anti- LPO IgY with ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin, casein and lysozyme were evaluated, resulting that the anti-LPO IgY found to be a specific antibody toward LPO and no cross-reaction was observed against ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin, casein, and lysozyme in double immunodiffusion test and ELISA test. In indirect ELISA method, coating concentration of LPO and dilution rate of anti-LPO IgY was 0.25$\mu\textrm{g}$/mL and 1:8,000 respectively. Sensitivity in the standard curve of LPO was ranged from 0.01 to 1$\mu\textrm{g}$/mL using anti-LPO IgY.

Development of a Blocking ELISA for Measuring Rabies Virus-specific Antibodies in Animals

  • Yang, Dong-Kun;Kim, Ha-Hyun;Ryu, Jieun;Gee, Mi-ryun;Cho, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.269-276
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    • 2018
  • Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

Development of a Quantitative ELISA for Anti HER-2 Antibodies using Human HER-2 Recombinant Proteins (인간 HER-2 재조합 단백질을 사용한 항 HER-2 항체 단백질의 ELISA 정량 방법 개발)

  • Jung, Sun-Ki;Ryu, Chang-Seon;Choung, Kyu-Jin;Song, Gyu-Yong;Kim, Sang-Kyum
    • YAKHAK HOEJI
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    • v.55 no.1
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    • pp.16-21
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    • 2011
  • HER-2 (Human Epidermal Growth Factor Receptor-2) is a protein giving higher aggressiveness in human breast cancers. Trastuzumab is a monoclonal antibody that targets HER-2 and is known to extend survival across all stages of HER2-positive breast cancer. In this study, we attempted to development of a quantitative ELISA (Enzyme-Linked ImmunoSorbent Assay) for evaluating anti HER-2 antibodies using human HER-2 recombinant proteins to support antibody producing processes and pharmacokinetic studies. We established direct or indirect ELISA method for the trastuzumab-like protein combined human recombinant HER-2. The ELISA method will prove to be great value in quantitating anti-HER-2 antibodies levels for developing anticancer antibodies.

The comparative study on Newcastle disease virus antibody titer by hemagglutination inhibition test and enzyme-linked immunosorbent assay (혈구응집억제반응과 효소면역측정법을 이용한 닭 뉴캣슬병 바이러스에 대한 혈중항체가 비교)

  • Han, Sung-Tae;Lee, Cheong-San;Kwak, Hak-Koo;Song, Jong-Han;Lee, Jong-In
    • Korean Journal of Veterinary Service
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    • v.26 no.3
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    • pp.215-219
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    • 2003
  • This study was conducted to investigate the similarity between hemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay(ELISA) titers and sample to positive ratio (S/P ratio) of Newcastle disease(ND) virus. To perform this study, the 372 sera of broiler chicks and 120 sera of layers and breed chicks were collected from slaughter house and farms, respectively. As a result of HI test out of different chicks, the positive percentage of ND antibody titer of broiler, layer and breeder, when a standard positive HI titer were '2', was 84.4%, 100% and 100%, respectively. The positive percentage of ND antibody titer by ELISA was shown 38.4%, 100% and 100% and S/P ratio were also shown 81.5%, 98.2% and 99.2%, respectively. The results of comparative survey with same sera by two experimental methods were as follows; In low HI titer, ELISA titer was not similar to HI titer, but S/P ratio was similar to it. In high HI titer, ELISA titer was not similar to HI titer. Therefore, HI titer was more similar to S/P ratio than ELISA titer.

Production and Characterization of a Monoclonal Antibody Against Enrofloxacin

  • Chusri, Manaspong;Wongphanit, Pitikarn;Palaga, Tanapat;Puthong, Songchan;Sooksai, Sarintip;Komolpis, Kittinan
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.69-75
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    • 2013
  • Enrofloxacin is a fluoroquinolone antibiotic approved for the treatment of infections in animals. Because of the side effects to consumers of animal products, the maximum residue limits (MRLs) of enrofloxacin in animal tissues for consumption are regulated. In this study, a monoclonal antibody (mAb) against enrofloxacin was prepared and characterized for the development of a direct competitive enzyme-linked immunosorbent assay (ELISA). The obtained mAb, Enro44, was highly specific for enrofloxacin and had a 50% inhibition concentration ($IC_{50}$) of 1.99 ng/ml in a competitive ELISA, and the limit of detection (LOD) was 0.50 ng/ml. The cross-reactivity of the mAb with other quinolones and fluoroquinolones was lower than 0.01%. The subclass of the mAb Enro44 was identified as IgG1. The antigen (Ag)-captured direct competitive ELISA using the mAb Enro44 was tested on different spiked samples, including chicken muscle, cattle milk, and cattle urine, and the assay demonstrated recoveries of 82-112%, 80-125%, and 78-124%, respectively. Furthermore, the quantitation of enrofloxacin obtained from the ELISA and from high-performance liquid chromatography (HPLC) was in good agreement, with the linear regression coefficient between 0.933 and 1.056. The cDNAs encoding a heavy-chain Fd fragment (VH and CH1) and a light chain of the mAb Enro44 were cloned and sequenced. Taken together, the results obtained reveal a potential use of this mAb in an ELISA for the detection of enrofloxacin in food samples. The information of amino acid sequence of this mAb will be useful for further modification and production of the mAb in a bioreactor.

Monoclonal Antibody-Based Dipstick Assay: A Reliable Field Applicable Technique for Diagnosis of Schistosoma mansoni Infection Using Human Serum and Urine Samples

  • Demerdash, Zeinab;Mohamed, Salwa;Hendawy, Mohamed;Rabia, Ibrahim;Attia, Mohy;Shaker, Zeinab;Diab, Tarek M.
    • Parasites, Hosts and Diseases
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    • v.51 no.1
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    • pp.93-98
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    • 2013
  • A field applicable diagnostic technique, the dipstick assay, was evaluated for its sensitivity and specificity in diagnosing human Schistosoma mansoni infection. A monoclonal antibody (mAb) against S. mansoni adult worm tegumental antigen (AWTA) was employed in dipstick and sandwich ELISA for detection of circulating schistosome antigen (CSA) in both serum and urine samples. Based on clinical and parasitological examinations, 60 S. mansoni-infected patients, 30 patients infected with parasites other than schistosomiasis, and 30 uninfected healthy individuals were selected. The sensitivity and specificity of dipstick assay in urine samples were 86.7% and 90.0%, respectively, compared to 90.0% sensitivity and 91.7% specificity of sandwich ELISA. In serum samples, the sensitivity and specificity were 88.3% and 91.7% for dipstick assay vs. 91.7% and 95.0% for sandwich ELISA, respectively. The diagnostic efficacy of dipstick assay in urine and serum samples was 88.3% and 90.0%, while it was 90.8% and 93.3% for sandwich ELISA, respectively. The diagnostic indices of dipstick assay and ELISA either in serum or in urine were statistically comparable (P>0.05). In conclusion, the dipstick assay offers an alternative simple, rapid, non-invasive technique in detecting CSA or complement to stool examinations especially in field studies.