• Journal of Applied Biological Chemistry
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• v.62 no.3
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• pp.287-292
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• 2019

Pseudomonas tolaasii causes brown blotch disease on the oyster mushroom (Pleurotus ostreatus). Various pathogenic strains of P. tolaasii were isolated and divided into three subtypes, $P1{\alpha}$, $P1{\beta}$, and $P1{\gamma}$. For phage therapy, bacteriophages against to these subtype strains were applied to mushroom cultivation and very successful to prevent from the disease. In this study, bacteriophages were isolated against the representative strains of subtype pathogens and their polyclonal antibodies were synthesized to investigate structural relationship among capsid proteins of phages. Phage preparations over $10^{10}pfu/mL$ were injected to rabbit thigh muscle and polyclonal antibodies were obtained after three times of boost injection. Titers of the antibodies obtained were over $2{\times}10^7Ab/mL$ for the phage ${\phi}6264$, $1{\times}10^6Ab/mL$ for the phage ${\phi}HK2$, and $1{\times}10^7Ab/mL$ for the phage ${\phi}HK19$ and phage ${\phi}HK23$. High specific activities were observed between antibodies and the corresponding bacteriophages. Some cross-reactivities between the antibodies and non-corresponding bacteriophages were also measured. Antibody $Ab{\phi}6264$ inactivated all phages of $P1{\alpha}$ subtype and only phage ${\phi}HK16$ among $P1{\beta}$ subtype phages. Antibody $Ab{\phi}HK23$ of $P1{\gamma}$ subtype neutralized all phages of $P1{\beta}$ subtype as well as the phage ${\phi}HK23$, showing the widest phage-inactivation range. When the structural-similarity studies of phages were investigated by using phage antibodies, closeness obtained by phylogenetic analysis of 16S rRNA genes of pathogenic strains were quite different from that of polyclonal antibody-specific structural similarity of phage capsid proteins. In conclusion, there is weak correlation between the host strain specificity of bacteriophage and its capsid structural similarity measured by phage antibodies.

• Journal of Animal Science and Technology
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• v.47 no.1
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• pp.19-28
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• 2005

The objectives of the current study were to develop polyclonal antibodies in sheep against adipocyte plasma membrane(APM) proteins isolated from swine, to investigate tissue specificity, and to determine cytotoxic effects of antiserum on swine adipocytes. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver, and spleen were isolated using a 32% sucrose gradient. Adult male sheep was immunized three times at three week interval with the purified swine APM proteins. Antiserum was taken from immunized sheep at 10, 12, and 14 days after the third immunization. Antiserum expressed strong reactivity with APM proteins determined by enzyme-linked immunosorbent assay(ELISA), and the reactivity could be detected at dilutions in excess of 1 : 81,000. Antiserum showed very low binding affinity with proteins isolated from brain, heart, kidney, liver, or spleen. Tissue specificity of the antiserum was reconfirmed by Western immunoblotting using anti-sheep immunoglobulin G•alkalinephosphatase conjugate as a secondary antibody. The reactivity of antiserum to the external surface of fixed swine adipocytes was confmned by an immunohistochemical technique using anti-sheep immunoglobulin G-FITC. Confluent swine adipocytes in culture were lysed by antiserum treatment and cytosolie lactate dehydrogenase(LDH) was released as a dose-dependent patterns while adipocytes treated with normal sheep serum maintained their integrity and expressed low level of LDH. These results implicate that fat contents in the pigs can be reduced by immunological methods.

• Parasites, Hosts and Diseases
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• v.60 no.2
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• pp.143-147
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• 2022

Acanthamoeba keratitis (AK) is a rare ocular disease, but it is a painful and sight-threatening infectious disease. Early diagnosis and adequate treatment are necessary to prevent serious complications. While AK is frequently diagnosis via several PCR assays or Acanthamoeba-specific antibodies, a more specific and effective diagnostic method is required. This study described the production of a polyclonal peptide antibody against the periplasmic binding protein (PBP) of A. castellanii and investigated its diagnostic potential. Western blot analysis showed that the PBP antibody specifically reacted with the cell lysates of A. castellanii. However, the PBP antibody did not interact with human corneal epithelial (HCE) cells and the other 3 major causative agents of keratitis. Immunocytochemistry (ICC) results revealed the specific detection of A. castellanii trophozoites and cysts by PBP antibodies when A. castellanii were co-cultured with HCE cells. PBP antibody specificity was further confirmed by co-culture of A. castellanii trophozoites with F. solani, S. aureus, and P. aeruginosa via ICC. The PBP antibody specifically reacted with the trophozoites and cysts of A. polyphaga, A. hatchetti, A. culbertsoni, A. royreba, and A. healyi, thus demonstrated its genus-specific nature. These results showed that the PBP polyclonal peptide antibody of A. castellanii could specifically detect several species of Acanthamoeba, contributing to the development of an effective antibody-based AK diagnostics.

• Journal of Applied Biological Chemistry
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• v.52 no.4
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• pp.170-173
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• 2009

Vitellogenin (Vtg) is found in the serum of both female and male fish that have been exposed to environmental endocrine disrupters or estrogen hormones, and is used as a biomarker for such contamination. In our current study antibodies raised against the purified flatfish Vtg were tested for their reactivity on immunoblots to flatfish and carp Vtg, and also to a Vtg protein fragment produced in E. coli. Polyclonal antibodies raised against purified flatfish Vtg reacted well with Vtg in the serum of flatfish and carp induced with $17{\beta}$-estradiol, but not with the Vtg protein fragment produced in E. coli.

• Food Science and Biotechnology
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• v.15 no.6
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• pp.990-992
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• 2006

In a previous study, we obtained polyclonal antibodies against the organophosphorus insecticide fenitrothion and developed an enzyme-linked immunosorbent assay (ELISA) for this pesticide. Using these antibodies and an enzyme tracer, a direct competitive ELISA method specific for fenitrothion using a dipstick format was developed. Dipstick ELISA using antibodies to fenitrothion immobilized on an Immunodyne membrane allowed the quick visual detection of fenitrothion at concentrations above $10\;{\mu}g/L$. The $IC_{50}$ value of dipstick ELISA using reflectance detection was $27\;{\mu}g/L$ with a detection limit of $2\;{\mu}g/L$. The recovery of fenitrothion from spiked lettuce and rice samples using the dipstick ELISA ranged from 87-107%.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.34 no.s01
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• pp.48-54
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• 1989

An immunoassay techniques using monoclonal antibodies have been developed of the quantitative analysis of endogenous hormones in plants. In order to be useful for immunoassay, both a high degree of specificity and a high affinity are required. A system is described for production of hybridomas which secrete antibodies against the plant hormone. Using the system we were able to produce hybridmas with the desired antibody specificity by cell fusion and culture method. For a number of obvious reasons, monoclonal antibodies(mAb) were superior to polyclonal antibodies.

• Journal of fish pathology
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• v.24 no.2
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• pp.75-84
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• 2011

The amino acid sequence of glycoprotein of Korean VHSV isolate (KR'01-1) was analyzed using the DNAStar Protean system. Based on the flexibility, hydrophilicity, antigenic index and surface probability, three regions (Gp1, Gp2 and Gp3) were selected as potential antigenic determinants. Three oligopeptides containing the amino acid sequences of the three regions were synthesized and polyclonal antibodies were raised against them. The activities of the antibodies were analyzed by Western blotting and virus neutralization test. The results showed that antibodies raised against oligopeptides Gp1 and Gp2 neutralized the infectivity of VHSV, suggesting that they can be possible candidates for subunit vaccines against VHS diseases in olive flounder.

• Biomolecules & Therapeutics
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• v.2 no.4
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• pp.303-309
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• 1994

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. Two kinds of antibodies could be produced, one is from synthetic peptides and the other from proteins such as purified receptor. Anti-peptide antibodies gave some advantages; epitope is evident and also receptor purification in quantity is not prerequisite. It can be also applied to the study of receptor structure-activity relationship. The purpose of the present study was 1) to produce and characterize a polyclonal antibody against a synthetic $\beta$2-adrenergic receptor peptide(Phe-Gly-Asn-Phe-Trp-Cys-Phe-Trp-Thr-Ser-Ile-Asp-Val-Leu) and 2) to determine the effects of this antibody on the $\beta$-adrenergic receptor ligand interaction. The peptide sequence contains an amino acid residue such as Asp-113 which was identified as one of important component for receptor-ligand interaction in site-directed mutagenesis studies. Production of antibody was performed by immunization of rabbits through popliteal lymph node with the peptide coupled with Keyhole Limpet Hemocyanin (KLH). The titer of antibody against this peptide was 1 : 1000. The anti-peptide antibody was able to detect a 67 kDa protein band in western blot corresponding to the molecular weight of the $\beta$-adrenergic receptor in partially purified receptor fraction derived from guinea pig lung. The antisera inhibited the specific binding of [$^3$H]dihydroalprenolol to $\beta$-adrenergic receptor in a concentration-dependent manner. The results from this study suggest that the peptide sequence selected in the present study is important for the receptor ligand interaction.

• Korean Journal of Food Science and Technology
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• v.46 no.5
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• pp.538-543
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• 2014

A sandwich enzyme-linked immunosorbent assay method (sELISA) for detecting the presence of shrimp in processed foods was developed using rabbit polyclonal antibodies against tropomyosin produced by black tiger prawns (shrimp). Based on the standard curve derived using this method, the detection range of shrimp was determined to be $1-100{\mu}g/mL$. The cross-reactivity of these antibodies toward black tiger prawns, fleshy prawns, cocktail prawns, lobster, and blue crab was 100, 73, 155, 18, and 0%, respectively. When shrimp was heated for 10 min, the mean assay recovery of tropomyosin was 121-221% at $70-100^{\circ}C$ and 7.8% at $121^{\circ}C$. When shrimp was added to cream soup, weaning food, sausage, fish paste, and sauce, the mean assay recovery was 397, 639, 168, 234, and 0%, respectively. In sample tests involving 14 commercial items, the coincidence ratio of assay results and reference was 79%.

• BMB Reports
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• v.29 no.5
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• pp.462-467
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• 1996

Acetolactate synthase (ALS, EC 4.1.3.18) is the first common enzyme in the biosynthesis of leucine, isoleucine, and valine. It is the target enzyme for several classes of herbicides, including the sulfonylureas, the imidazolinones, the mazolopyrimidines, the pyrimidyl-oxy-benzoates, the pyrimidyl-thio-benzens, and the 4,6-dimethoxypyrimidines. An amino-terminal fragment of the sulfonylurea-resistant ALS gene (SurB) from Nicotiana tabaccum was cloned into the bacterial expression vector pGEX-2T. The resulting recombinant plasmid pGEX-ALS1 was used to transform Escherichia coli strain BL21, and the tobacco ALS was expressed in the bacteria as a protein fused with glutathione S-transferase (GST). Polyclonal antibodies against the fusion product (GST-ALS) were produced, and the sensitivity of GST-ALS with the rabbit anti-GST-ALS IgG was up to 50 ng. This antibody was used for Western blot analysis of the partially purified ALS from barley shoots. The results suggest that the polyclonal antibody produced in this study can be used to detect plant ALS.