• Korean Journal of Animal Reproduction
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• v.14 no.2
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• pp.125-131
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• 1990

These experiments were carried out to investigate of the molecular characteristics of porcine zona pellucidae and to examine the reactivity of anti-zona antibody by SDS-PAGE, Immunoblotting and Immunoprecipitation. The results obtained in these experiments were summarized as follows : 1. The result obtained by SDS-PAGE of porcine zona pellucidae indicated that it composed of several units with molecular weight ranging 55,000-110,000. 2. In order to see the reactivity of antibodies to zona pellucidas, immunoblotting was applied. The results indicated that polyclonal antibodies to porcine and mouse zona reacted with porcine zona. While monoclonal antibody to porcine did not react with the procine zona enough to show a clear band on a gel. 3. Labelling of porcine zonae with 125I was performed using the Iodogen method, the radioactivity and the percent incorporation of 125I into porcine zonae were approximately 26,000 cpm/10${mu}ell$ and 16, respectively. 4. Measurements of radioactivity and O.D value for Immunoprecipitates produced by reaction of 125I-porcine zona with anti-zona antisera were confirmed that existence of reactivity of monoclonal antibody to porcine zona although its reactivity was lower than that of polyclonal antibodies, and reconfirmed that cross-reactivity of polyclonal antibody of mouse zona with porcine zona.

• The Plant Pathology Journal
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• v.32 no.5
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• pp.452-459
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• 2016

The genomic region of Grapevine fanleaf virus (GFLV) encoding the movement protein (MP) was cloned into pET21a and transformed into Escherichia coli strain BL21 (DE3) to express the protein. Induction was made with a wide range of isopropyl-${\beta}$-D-thiogalactopyranoside (IPTG) concentrations (1, 1.5, and 2 mM) each for duration of 4, 6, or 16 h. However, the highest expression level was achieved with 1 mM IPTG for 4 h. Identity of the expressed protein was confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting. The expressed 41 kDa protein was purified under denaturing condition by affinity chromatography, reconfirmed by Western blotting and plate-trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA) before being used as a recombinant antigen to raise polyclonal antibodies in rabbits. Purified anti-GFLV MP immunoglobulines (IgGs) and conjugated IgGs detected the expressed MP and GFLV virions in infected grapevines when used in PTA-ELISA, double antibody sandwich-ELISA, and Western blotting. This is the first report on the production of anti-GFLV MP polyclonal antibodies and application for the virus detection.

• Journal of Microbiology
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• v.33 no.3
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• pp.201-207
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• 1995

We have previously reported that SPP2 gene product of yeast Saccharomyces cerevisiae is involved in the pre-mRNA splicing. To investigate the rol ein the splicing pathway of the Spp2p protein, the SPP2 gene was overexpressed in Escherichia coli and polyclonal antibodies to Spp2p were generated from rabbits. First, a DNA fragment containing the SPP2 GENE without its promoter was subcloned into an E. coli expression vector, pKK233-3. The resulting recombinant plasmid pBQ14 contained an IPTG inducible tac promoter and the SPP2 structural gene. Overexpression of the SPP2 gene was achieved by additionof 0.1 to 1.0 mM IPTG to a logarithmic culture of E. coli JM103(pBQ14) for 90 min at 37.deg.C. Sequence of N-terminal 15 amino acids of the overproduced protein was well matched to the deduced one from the SPP2 reading frame. Then, polyclonal antibodies were generated from rabbits immunized with gel-purified SppSp protein. These antibodies reacted specifically with the Spp2p protein extracted from yeast cells expressing the SPP2 gene to a great extent. The antibodies could also block the activity of yeast splicing extracts.

• Journal of Life Science
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• v.19 no.2
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• pp.284-288
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• 2009

The innate immune system is important for the first line of host defence against infectious agents, which have penetrated the mechanical barriers. Mannose-binding lectin (MBL or mannan-binding protein, MBP) is a serum protein that is synthesized in the liver as a part of the acute phase response. MBL binds to carbohydrate structures presented by a wide range of pathogenic bacteria, viruses, fungi, and parasites. MBL is synthesized as a monomer that has a carboxy-terminal carbohydrate recognition domain, a neck region and a collagen region. Low MBL level was reported to be the most frequent immuno-deficiency syndrome. Although extensive studies have yielded detailed information on the structure of MBL, functions of the MBL complex are not fully understood yet. We, here, present cloning process of MBL cDNA from the rat liver and production of truncated recombinant MBL protein using a bacterial expression system in order to produce anti-MBL polyclonal antibody. Anti-MBL polyclonal antibody was raised in a New Zealand rabbit and its affinity was tested against recombinant protein using western blot technique. MBL cDNA, recombinant protein and anti-MBL antibody could be used as great arsenals to dissect cellular biochemistry of MBL.

• Food Science and Biotechnology
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• v.16 no.4
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• pp.515-519
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• 2007

Rapid immunochromatographic assay (ICA) kits were developed using flagella-specific monoclonal antibodies (MAbs) and rabbit polyclonal antibodies for screening Listeria spp. in food. The establishment of different formats, MAb 2B1 as capture antibody and MAb 7A3 or rabbit polyclonal antibodies as detector antibody, was compared. The 2 formats of the ICA kit were shown to have specific reactions with Listeria and no cross-reactivity with any of the non-Listeria including Escherichia coli O157:H7 and Salmonella enteritidis. The detection limits of the ICA kit using the combination of gold-labeled MAb 7A3 and MAb 2B1 showed $1{\times}10^5$ and $1{\times}10^6\;CFU/0.1\;mL$ at 22 and $30^{\circ}C$, respectively. The other format of the ICA kit using the combination of gold-labeled rabbit polyclonal antibodies and MAb 2B1 showed $1{\times}10^6\;CFU/0.1\;mL$ at $22^{\circ}C$ but weak signal at 30 culture. The format utilizing MAb was more sensitive than the one using polyclonal antibodies for capture antibody. Samples contaminated with L. monocytogenes 4b culture (9-10, 5-6, and 1-2 CFU/mL) on pork and pasteurized milk were confirmed as positive results. Current data suggests that this ICA kit is a rapid, simple and effective tool to screen for Listeria spp. in food.

• The Korean Journal of Pesticide Science
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• v.5 no.2
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• pp.1-12
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• 2001

A competitive indirect enzyme-linked immunosorbent assay (ci ELISA) for the organophosphorus insecticide acephate, O,S-dimethyl acetylphosphoramidothioate, was developed using a polyclonal antibody. Three different haptens mimicking the analyze and containing hexanoic acid moiety as a linker were synthesized, and then conjugated with the carrier proteins bovine serum albumin and keyhole limpet hemocyanin by the N-hydroxysuccinimide active ester method. Polyclonal antibodies raised against hapten-KLH conjugates in rabbits and the hapten-BSA conjugates as coating antigens were screened and selected for the assay in the homologous and/or heterologous ELISA system. The effects of various assay conditions, including blocking reagents, detergent content, organic solvents, pH, and preincubation of tile mixture of the polyclonal antibody and the analyze on the sensitivity were evaluated. The $IC_{50}$ value of acephate of 110 ng/mL was obtained in an optimized heterologous system using hapten-3-BSA as a coating antigen and a polyclonal antibody 8377, showing the detection range of 10-1000 ng/mL and the lowest detection limit of 4 ng/mL. The cross-reactivities of the structurally related insecticides, including methamidophos were less than 0.02%. These results indicate that the ELISA could be a convenient and alternative tool for monitoring acephate residues in agricultural products and environmental samples.

• 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.

• Journal of Korean Neurosurgical Society
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• v.30 no.6
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• pp.774-776
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• 2001

We report a 35-year old female patient with history of seizure and mass which was confirmed as a plasma cell granuloma, arising in the left parietal area. The mass appeared on magnetic resonance imaging as well circumscribed area of decreased signal that markedly enhanced with administration of the contrast. Pathologically, biopsy showed a mixed cellular population with considerable numbers of plasma cells along with eosinophils and lymphocytes and the tumors was characterized immunohistochemically by polyclonal population of lymphoid cells.

• BMB Reports
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• v.31 no.3
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• pp.240-244
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• 1998

Helicobacter pylori is the etiologic agent of human gastritis and peptic ulceration and produces urease as the major protein component on its surface. H. pylori urease is known to serve as a major virulence factor and in a potent immunogen. In order to express the recombinant urease at a higher level, a DNA fragment containing the minimal H. pylori urease gene cluster was subcloned into a high copy-number vector. The recombinant H. pylori urease expressed in an E. coli strain that was grown in a rich medium supplemented with added nickel was purified to near homogeneity by using DEAE-Sepharose, Superdex HR200, and Mono-Q (FPLC) columns and the purified enzyme possessed the specific activity of 1255 U/mg. Polyclonal antibodies raised against the purified recombinant H. pylori urease were shown to be very specific when subjected to Western blot analysis, in which crude extracts from the H. pylori ATCC strain and the recombinant E. coli strains expressing various bacterial ureases were exnmined for cross-reactivity.

• BMB Reports
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• v.30 no.6
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• pp.448-452
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• 1997

Polyclonal antibody of the boar sperminogen was used to characterize the boar sperm proteinase sperminogen. Boar sperminogen was purified from the acid extracts of the washed epididymal spermatozoa by gel filtration through a Sephadex G-100 column. followed by preparative SDS-PAGE. The sperminogen band was sliced out and was eluted from the gel matrix. The purified sperminogen was used to produce the polyclonal antibody of the boar sperminogen. When characterized on a Western blot, the final preparation of sperminogen appeared as a homogenous protein with a molecular weight of 32 kDa. The relative migration of sperminogen was distinctly different from the major components of the proacrosin-acrosin system as well as all the observable proacrosin activation by-products detected on the Western blot. The sperminogen antibody, however. cross-reacted with the proacrosin-acrosin system.