Yong-Seung Shin

Production of monoclonal antibodies against serum immunoglobulins of black rockfish (Sebastes schlegeli Higendorf).

The present study was undertaken to produce monoclonal antibodies (MAbs) against immunoglobulin (Ig) purified from black rockfish (Sebastes schlegeli Higendorf) serum using protein A, mannan binding protein, and goat IgG affinity columns. These three different ligands were found to possess high affinity for black rockfish serum Ig. All of the Igs purified eluted at only 0.46 M NaCl concentration in anion exchange column chromatography and consisted of two bands at 70 kDa and 25 kDa in SDS-PAGE; they also had similar antigenicity for MAbs to Ig heavy chain in immunoblot assays. Therefore, black rockfish Ig is believed to exist as a single isotype within serum. The MAbs produced against Ig heavy chain reacted specifically with spots distributed over the pI range from 4.8 to 5.6 with a molecular weight of 70 kDa on two dimensional gel electrophoresis immunoblot profiles.

Physiological Function of Insoluble Dietary Fiber Prepared from Exploded Oak Wood (Quercus mongolica)

This study investigated the production of insoluble dietary fiber using exploded and chemically treated oak wood (Quercus mongolica) and the physiological functions of prepared insoluble dietary fiber in laboratory animals. To produce high quality insoluble dietary fiber, the steam explosion treatment was performed at 25 kgf/cm2 pressure for 6 minutes. In the chemical analysis of insoluble dietary fiber, exploded oak wood was pretreated by 1% sodium hydroxide solution. The insoluble dietary fiber contained 7.6% residual lignin and 61.7% of alpha-cellulose. In order to compare the physiological functions of prepared insoluble dietary fiber with those of commercial insoluble dietary fiber, Sprague-Dawley male rats weighing 100 +/- 10 g were randomly assigned to one normal diet and five high cholesterol diets, containing 1% cholesterol. The high cholesterol diet groups were classified as the fiber-free diet (FF group), 5% commercial alpha-cellulose diet group (5C group), 10% commercial alpha-cellulose group (10C group), 5% insoluble dietary fiber group (5M group) and 10% insoluble dietary fiber group (10M group). Food intake, weight gain and food efficiency ratio in high cholesterol groups were significantly higher than those of the normal group, but there were no significant differences among the high cholesterol diet groups. In addition, there were no significant differences in the weights of liver, kidney and small intestine in insoluble dietary fiber-supplemented groups. Cecum weights in all insoluble dietary fiber groups were significantly higher than those of the FF group. There were no significant differences in the activities of the glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT) among the insoluble dietary fiber-supplemented groups. In conclusion, the prepared insoluble dietary fiber and the commercially available insoluble fiber showed the same physiological effects. Moreover, the preparation method for the insoluble dietary fiber from the exploded oak wood was successful.

Development of competitive ELISA for neosporosis by employing immunoproteomics

In this study, proteomics was used to explore the antigenic proteins that are involved in cross-reactivity during serodiagnosis between Neospora caninum (N. caninum) and Toxoplasma gondii (T. gondii). Competitive enzyme-linked immunosorbent assay (C-ELISA) developed by proteomics shed a new light on the infection of N. caninum. Cross-reactivity of antigenic proteins between N. caninum and T. gondii tachyzoites was explored by using the conventional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) (1-DE) and two-dimensional gel electrophoresis (2-DE) immunoblot. The proteins were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The protein expression patterns in the immunoblot profiles of N. caninum were similar to bovine, chicken, and rabbit anti-N. caninum serum, but they were not similar to rabbit anti-T. gondii serum. Band at 79 kDa, HSP70, and actin on immunoblot profiles reacted, in general, with bovine, chicken, and rabbit anti-N. caninum serum and also with rabbit anti-T. gondii serum, respectively. Whereas the band at 144 kDa, and NCDG-1 were detected on bovine, chicken, and rabbit anti-N. caninum immunoblot profiles, they were not observed on rabbit anti-T. gondii immunoblot profile. These specific antigenic proteins were recorded as species-specific proteins of N. caninum against T. gondii. Based on the proteome analysis, C-ELISA was developed to screen the cattle infected with N. caninum by using N. caninum tachyzoite lysate as a coating antigen and chicken anti-N. caninum immunoglobulin (Ig)Y as a competitor. C-ELISA was able to detect the antibody of N. caninum without cross-reactivity with T. gondii. Furthermore, it achieved a fine diagnostic performance in the cases of 162 bovine sera.