• Korean Journal of Veterinary Research
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• v.33 no.1
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• pp.119-123
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• 1993

The present study was intended to use the avidin-biotin-peroxidase-antiperoxidase complex (ABPAP) method for the identification of Mycobacterium bovis in the tissue sections of infected cattle. Antibodies and linksera for ABPAP procedure used in incubated order were rabbit anti-Mycobacterium polyvalent antibodies, goat anti-rabbit IgG, rabbit peroxidase-antiperoxidase complex, biotinyl-horse anti-rabbit IgG, and avidin-biotin-peroxidase complex. Where the bacterial antigen was localized by ABPAP, a dark brown deposit occurred in the cytoplasms of macrophages and Langerhans' giant cells of the granulomatous lesions. The method approved to be highly specific for the identification of the bacteria and allowed a precise localization of the bacterial antigen in infected cells.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1855-1862
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• 2016

Cronobacter sakazakii (C. sakazakii) is a foodborne pathogen, posing a high risk of disease to infants and immunocompromised individuals. In order to develop a quick, easy, and sensitive assay for detecting C. sakazakii, a rabbit anti-C. sakazakii immunoglobulin G (IgG) was developed using sonicated cell protein from C. sakazakii. The developed anti-C. sakazakii (IgG) was of good quality and purity, as well as species-specific. The developed rabbit anti-C. sakazakii IgG was attached to the surface of a sulforhodamine B-encapsulated liposome to form an immunoliposome. A test strip was then prepared by coating goat anti-rabbit IgG onto the control line and rabbit anti-C. sakazakii IgG onto the test line, respectively, of a plastic-backed nitrocellulose membrane. A purple color signal both on the test line and the control line indicated the presence of C. sakazakii in the sample, whereas purple color only on the control line indicated the absence of C. sakazakii in the sample. This immunochromatographic strip assay could produce results in 15 min with a limit of detection of $10^7CFU/ml$ in C. sakazakii culture. The immunochromatographic strip assay also showed very good specificity without cross-reactivity with other tested Cronobacter species. Based on these results, the developed immunochromatographic strip assay is efficient for the detection of C. sakazakii and has high potential for on-site detection.

• Clinical and Experimental Reproductive Medicine
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• v.17 no.1
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• pp.71-80
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• 1990

New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests.

• Journal of Sensor Science and Technology
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• v.30 no.5
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• pp.337-341
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• 2021

In this paper, we describe the fabrication of a paper-based enzyme-linked immunosorbent assay (ELISA) to detect polygalacturonase (PG), which is used as a biomarker to determine whether a plant is infected with a disease. The proposed paper-based ELISA can analyze the concentration of PG in a short time using a small sample compared to the traditional ELISA, which is generally performed using a well plate. To increase the resolution of the sensor, we optimized the dilution ratio of the HRP-conjugated goat anti-rabbit IgG antibody and the dilution ratio of the anti-PG and HRP-conjugated goat anti-rabbit IgG antibodies. Furthermore, for quantitative analysis of PG concentration, Delta RGB analysis was conducted to detect color changes in the sensing window displayed by the PG samples at various concentrations. Based on the experiment, the fabricated paper-based ELISA could measure at least 0.25 ㎍ of PG and the measurement range was 0.25-2 ㎍. Therefore, the paper-based ELISA for detecting PG is expected to be able to determine the presence or absence of disease in crops at the infection stage in the future.

• Korean Journal of Environmental Biology
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• v.19 no.4
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• pp.302-312
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• 2001

Immunoglobulin G was purified by 40% $(NH_4)_2SO_4$ precipitation, DEAE-Sephadex, Sephadex G-150 column chromatographies from rabbit antiserum against V. vulnificus ATCC 27562 O antigen and used for immunological test for V. vulnificus isolates. The profiles of cell lysate total protein and outer membrane protein from the isolates were analyzed by SDS-PAGE and densitometry. The overall profiles in all isolates were similar. Distict protein band was observed in comparison with V. parahaemolyticus. Western Blotting with rabbit Immunoglobulin G against cell lysates and OMP of V. vulnificus isolates showed a strong antigenic response to antigen 66, 60, 54, 48, 33 and 26 kDa which were common to all strains examined. The 26 kDa antigen showed V. vulnificus specific antigen in comparison with Vibrio parahaemolyticus. A sandwich enzyme-linked immunosorbent assay was developed by using rat anti-V. vulnificus ATCC 27562 polyclonal antibodies as capture antibody, a purified rabbit IgG antibody as detector antibody, and goat anti-rabbit IgG-alkaline phosphatase conjugate as developer antibody. When four V. vulnificus isolates were tested, the reactivity showed from 50 to 70% by sandwich ELISA.

• Journal of Microbiology and Biotechnology
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• v.23 no.4
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• pp.527-533
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• 2013

Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was $10^5$ CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail.

• Korean Journal of Veterinary Research
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• v.30 no.2
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• pp.215-221
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• 1990

The present study was intended to use the peroxidase-antiperoxidase method for the identification of hog cholera virus(HCV) in the lymphatic organs of HCV-infected pigs. Sections were incubated with primary antibody (rabbit anti-HCV polyclonal or mouse anti-HCV monoclonal), followed by incubation with linkserum (goat anti-rabbit IgG) in excess and rabbit or mouse PAP complex. The viral antigen was localized mainly in the cytoplasms of lymphoid cells and macrophages. Positive reaction cells were frequently detected in the marginal areas of the germinal centers of the spleens, and also found in the tensils and lymph nodes. The method approved to be highly specific for the identification of the virus and allowed a precise localization of the viral antigen in infected cells.

• Korean Journal of Veterinary Service
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• v.15 no.2
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• pp.184-194
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• 1992

This study was done to identify Newcastle disease virus(NDV) antigens in paraffin sections of various organs from experimentally NDV-infected chicken using peroxidase-antiperoxidase(PAP) technique. Sections were Incubated with rabbit anti-NDV polyclonal as first antibody, followed by incubation with goat anti-rabbit IgG conjugate and peroxidase anti-peroxidase ( PAP ). Positive reactions were often detected in the epithelim of trachea and in the lymphocyte of spleen at 24 hours after virus inoculation. The viral antigen was localized mainly in the cytoplasm of infected cells. The method approved to be highly specific for the indetification of NDV and allowed a precise localization of the viral antigens in infected cells.

• Parasites, Hosts and Diseases
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• v.33 no.3
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• pp.155-164
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• 1995

The location of actin and myosin of the several stages of Cwptosporinium parvum was observed. The tissue antigen of C. pcruum was prepared through immunosuppression of IgG mice with Depomedrol . The thin sectioned specimens, which were incubated with the IgG fraction of the rabbit polyclonal antibodies raised against chicken back muscle actin and bovine uterus myosin, were treated with 10 nm gold-conjugated goat anti-rabbit IgG, Electrodense particles were located mainly on the pellicles of all observed developmental stages of the parasites. The number of actin gold particles in the cytoplasm increased when the parasite was dividing actively as in case of meronts. Especially in macrogametocytes, a lot of actin and myosin particles were synthesized and storaged as amilopectin-like bodies. There were many actin gold particles along the microspikes of cytoplasmic membranes in various developmental stages. The actin and myosin observed in this study may play important roles to control the shape of the parasites and movement of cytoplasmic membranes as cvtoskeletal proteins.

• Food Science and Biotechnology
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• v.17 no.3
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• pp.547-552
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• 2008

The detection of carbamate (carbofuran, carbaryl, benfracarb, thiodicarb, and methomil) and organophosphate (diazinon, cadusafos, ethoprofos, parathion-methyl, and chlorpyrifos) pesticide residues with very low detection limits was carried out using surface plasmon resonance (SPR) based equipment. The capacity to develop a portable SPR biosensor for food safety was also investigated. The applied ligand for the immunoassays was polyclonal goat anti-rabbit immunoglobulin (IgG) peroxidase conjugate. Concentration tests using direct binding assays showed the possibility of quantitative analysis. For ligand fishing to find a proper antibody to respond to each pesticide, acetylcholinesterase (AChE), and glutathione-S-transferase (GST) were tested. The reproducibility and precision of SPR measurements were evaluated. With this approach, the limit of detection for pesticide residues was 1 ng/mL and analysis took less than 11 min. Thus, it was demonstrated that detecting multi-class pesticide residues using SPR and IgG antibodies provides enough sensitivity and speed for use in portable SPR biosensors.