• Food Science and Biotechnology
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• v.14 no.3
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• pp.410-412
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• 2005

Vibrio parahaemolyticus is a gram-negative bacterium which acts as a causative agent for food poisoning. Studies with respect to specific extracellular proteins of V. parahaemolyticus would be useful for the development of specific detection methods against V. parahaemolyticus. In our present study, outer membrane proteins (OMPs) of V. parahaemolyticus were obtained from insoluble traction of 1% sarkosyl treated-cell wall materials. SDS-PAGE analysis showed the presence of several conserved outer membrane proteins among five strains of V. parahaemolyticus, and three bands were identified as V. parahaemolyticus OMPs through MALDI-TOF analysis. Polyclonal antibodies enriched with anti-OmpU were obtained from immunized rabbits. The antibodies against these proteins may be useful for the development of detection methods for V. parahaemolyticus.

• 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.

• 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.

• Food Science of Animal Resources
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• v.30 no.4
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• pp.560-567
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• 2010

This study was conducted to evaluate the in vivo gastrointestinal survival and adhesive properties of orally administered Lactobacillus brevis FSB-1. ELISA conducted using polyclonal antibodies specific for L. brevis FSB-1 was able to detect the organism in feces; therefore, we used ELISA to determine the concentration of lactic acid bacteria in feces collected from Wister rats that had been administered $10^{10}$ cells/rat/d orally for 20 d. The mean recovery of L. brevis FSB-1 was approximately $10^{7.22}$ cells/g of wet feces during the oral administration period, and $10^{7.50}$ and $10^{7.46}$ at 8 and 10 d after the end of oral administration, respectively. These results indicate that L. brevis FSB-1 was able to survive in the gastrointestinal tract of rats, and that it had a high adhesive property in rat colons.

• Biomedical Science Letters
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• v.7 no.4
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• pp.161-166
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• 2001

Most mammalian cells take up glucose by passive transport proteins in the plasma membranes. The best known of these proteins is the human erythrocyte glucose transporter, GLUT1. High levels of heterologous expression far the transporter are necessary for the investigation of its three-dimensional structure by crystallization. To achieve this, the baculovirus expression system has become popular choice. However, Spodoptera frugiperda Clone 9 (Sf9) cells, which are commonly employed as the host permissive cell line to support baculovirus replication and protein synthesis, grow well on TC-100 medium that contains 0.1% D-glucose as the major carbon source, suggesting the presence of endogenous glucose transporters. Furthermore, very little is known of the endogenous transporters properties of Sf9 cells. Therefore, human GLUT1 antibodies would play an important role for characterization of the GLUT1 expressed in insect cell. However, the successful use of such antibodies for characterization of GLUT1 expression m insect cells relies upon their specificity for the human protein and lack of cross-reaction with endogenous transporters. It is therefore important to determine the potential cross-reactivity of the antibodies with the endogenous insect cell glucose transporters. In the present study, the potential cross-reactivity of the human GLUT1 antibodies with the endogenous insect cell glucose transporters was examined by Western blotting. Neither the antibodies against intact GLUT1 nor those against the C-terminus labelled any band migrating in the region expected fur a protein of M$_r$ comparable to GLUT1, whereas these antibodies specifically recognized the human GLUT1. Specificity of the human GLUT1 antibodies tested was also shown by cross-reaction with the GLUT1 expressed in insect cells. In addition, the insect cell glucose transporter was found to have very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.

• Journal of Food Hygiene and Safety
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• v.31 no.6
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• pp.445-450
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• 2016

Thermal-stable soluble proteins (TSSP) in livestock products has been recently reported. Therefore, the development of antibodies and immunoassay using a TSSP is useful because the presence of TSSP can be measured on processed food. In this study, the existence of TSSPs in pork fat and meat was confirmed and their antigenicity was investigated. The extracts from pork fat and meat by heating method were analyzed by SDS-PAGE with 5% stacking and 12% separating gels. The protein profiles from the raw pork fat and meat extracts (major band ranged 25 to 100 kDa) without cooking and heating treatments were significantly different compared to those from cooked and heated pork fat and meat extracts (several major bands > 100 kDa and < 30 kDa). This meant that non thermal-stable soluble proteins ranged from 25 to 100 kDa may be denaturated to insoluble proteins by cooking and heating treatments, and TSSPs were in pork fat and meat at kept their properties. The confirmed TSSPs were used as an immunogen to investigate their antigenicity. Eight mice (5 mice for pork fat and 3 mice for pork meat) were separately immunized with the TSSPs of pork fat and meat, and the anti-sera obtained from the immunized mice showed high titer values. Polyclonal antibodies against each target protein showed the specific reaction to pork fat and meat, individually. These indicated that TSSP could be used as an immunogen to produce antibodies such as monoclonal and polyclonal antibodies. In addition, antibodies specific to TSSP from pork fat and meat may be used as a bio-receptor in immunoassays for the identification of fraudulent adulteration with pork fat and meat in livestock products.

• Development and Reproduction
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• v.9 no.2
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• pp.135-142
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• 2005

Vitellogenins(VTGs) are the precursor of egg-yolk proteins in most oviparous species from invertebrates to vertebrates. In oviparose vertebrates, VTGs are synthesized in the liver and transported through the blood to oocytes. In female fish, concentrations of plasma VTG increase rapidly at onset of vitellogenesis in the normal reproductive cycle. Male fishes also possess the gene for VTG, but plasma concentrations of the protein typically remain small, presumably due to low levels of endogenous estrogens. However, exposure of males to exogenous estrogenic mimics can result elevated. Therefore, the VTG in fish can be used as a useful biomarker for appropriate tools of endocrine disrupting compounds effects. In this studies, we prepared the test methods that can measure the plasma VTG level in the gobies that live in polluted area with mimic estrogen. For the purpose, we purified VTG of floating goby(Chaenogobius annularis) and prepared specific monoclonal and polyclonal antisera to yolk protein, then developed a sandwich competitive ELISA system for measurement of plasma VTG levels. Validation for the ELISA system using monoclonal and polyclonal antibodies against VTG was tested. The absorbance curve of serial dilutions of serum from vitellogenic female was paralleled to the standard curve of VTG, but normal male was not paralleled. The developed sandwich ELISA system was measured for VTG levels in plasma of common goby(Acanthogobius flaviman) and javeline goby(A. hasta) as well as in plasma of floating goby(C. annularis).

• Journal of Biosystems Engineering
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• v.37 no.6
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• pp.420-428
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• 2012

Purpose: Porcine proliferative enteropathy(PPE), caused by the obligate intracellular bacterium Lawsonia intracellularis, is a widely distributed disease throughout the world causing substantial economic loss. The bacterial pathogen invades the intestinal epithelial cells which causes hyperplasia of the infected cells and leads to the process of disease pathogenesis. For diagnosing PPE in a pig farm in earlier stage, a rapid diagnosing test equipment is needed for farmers. To test the equipment appropriately, we prepare the samples and antibodies for rapid detecting the Lawsonia intracellularis in pig feces. Methods : To prepare the PPE infected samples, we sampled PPE suspected pig feces in a pig farm. To manufacture a anti-Lawsonia intracellularis antibody for capturing the Lawsonia intracellularis, the rabbit-anti LsaA synthetic peptide polyclonal antibody was inoculated to rabbits. To select the couple of antibodies which is most well sandwiched with the bacteria, ELISA test was done with PPE infected ileum samples. Finally, to verify the PPE infected feces which would be used to test the rapid kit, PCR test was done on the sampled PPE suspected feces Results : The rabbit-anti LsaA synthetic peptide polyclonal antibody is developed, and is verified to capture the bacterial well through the fluorescence antibody test. Also, we found that the monoclonal antibody and the polyclonal antibody could be used as couples for sandwiching the bacteria. Finally, through the PCR test for samples of pig feces, we could prepare the 150 PPE positive samples and 50 PPE negative samples. Conclusions : The manufactured polyclonal antibody and the imported monoclonal antibody could be used to capture the bacteria using the sandwich techniques. Also, the prepared PPE infected negative and positive samples could be used to test the performance of the rapid kit to capture the bacterium Lawsonia intracellularis.

• Journal of Animal Science and Technology
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• v.51 no.3
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• pp.231-240
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• 2009

Sixteen ruminally cannulated Korean native steers (Hanwoo; $626.2\pm47.72$ kg) were used to investigate the effects of polyclonal antibodies against abdominal (AAb) and subcutaneous adipocyte membrane proteins (SAb) on ruminal fermentation patterns and blood metabolites. The body weight (BW) of Hanwoo was decreased 2-weeks after AAb and SAb injection, BW reduction was also observed in control and non-immunized serum groups, indicating that stress induced by other factors (e.g. blood sampling etc.) rather than antibodies injection may affect the BW reduction. Antibodies treatment did not affect (P > 0.05) rumen pH, volatile fatty acids and ammonia-N concentration. The ranges were similar with typical ranges of those in Hanwoo. Compared with control, blood urea N concentration was decreased in AAb group and increased (P < 0.05) in SAb group before antibodies treatment. However, none of the groups were significantly (P > 0.05) affected at 2- or 4-weeks after the treatment. Concentration of plasma glucose in the non-immunized serum group was significantly higher (P < 0.05) than the other groups at 0-week after treatment. However, antibodies treatment did not affect the concentration of plasma glucose. Concentration of plasma triglyceride showed no difference (P > 0.05) between the groups and ranged from 11.4 to 19.9 mg/dl, which is the perfect range of plasma triglyceride of Hanwoo fed concentrate based diets. In conclusion, these results may indicate that the present AAb and SAb have safety in nutritional physiological metabolism in Hanwoo. Further study on in vivo fat reduction of the antibodies against abdominal and subcutaneous adipocytes PMPs of Hanwoo is required for inedible fat-reduced high quality beef production.

• Journal of Animal Science and Technology
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• v.52 no.5
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• pp.375-382
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• 2010

Twelve pigs were used to investigate the effects of polyclonal antibody candidate against abdominal (AAb) and subcutaneous adipocyte membrane proteins (SAb) on body weight, fecaldigestibility and blood metabolites. When AAb and SAb developed by Choi et al. (2010) were injected to pigs, the numerical increase in BW (body weight) occurred at 4 weeks post-treatment, but BW for an entire period was also increased, indicating that the BW increase may not be affected by the antibodies injection. Antibodies treatment did not affect (P>0.05) fecal digestibility of dry matter, crude protein, crude fat and crude fiber. Fecal digestibility of crude ash for control (no treatment) at 2 weeks decreased, and that for non-immunized serum treatmentgroup at 4 weeks post-treatment increased, respectively (P<0.05). However, fecal digestibility of crude ash for AAb and SAb groups did not significantly change. At 4 weeks after the antibodies treatment, blood urea N concentration for AAb and SAb groups was significantly increased (P<0.05). However, these increases may not be caused by the antibodies treatment because similar pattern in blood urea N concentration occurred before the antibodies treatment. Antibodies treatment did not affect concentration of plasma glucose and triglycerides (P<0.05). Compared with control, concentration of plasma total cholesterol for AAb and SAb groups at 4 weeks post-treatment was significantly (P<0.05) decreased. This may suggest that body fat reduction possibly occurs. In conclusion, the AAb and the SAb developed by Choi et al. (2010) may have safety in nutritional physiological metabolism in pigs. Further study on in vivo fat reduction of the antibodies against abdominal and subcutaneous adipocytes of pigs should be required for fat-reduced pork production.