• Parasites, Hosts and Diseases
• /
• v.32 no.4
• /
• pp.249-258
• /
• 1994

Recently, the importance of toxoplasmosis is raised as a complication in immunosuppressed or AIDS patients. Our study focused on the identification of a variety of Toxoplasma antigens by immunoblotting. Rabbits and BALB/c mice were immunized with Toxoplosmo Iysate (RH strain) , frozen tachyzoites (RH strain) or cysts (Beverly and Fukaya strain) . Blood were collected from ear vein, heart or orbital plexus for detecting the serum antibody levels. For excretory-secretory (E.S) antigens, T gondii (RH) tachyzoite were cultured in CHL (Chinese hamster lung) cells with MEM containing of 5% FCS. After 72hrs, culture supernatant was collected. BALB/c mice were inoculated with RH tachyzoite intraperitoneally and peritoneal fluids were extracted three days later. E.S antigens were detected in culture supernatant and infected mouse peritoneal fluid by EITB. Serum IgG levels in rabbit were 1 :512 of 10 days after primary immunization, 1 : 2,048 of 10 days after secondary immunization, 1: 1,024 of 20 days after secondary immunization by IFAT, respectively. Serum IgG levels of immunized mice were 1:128 after 7 weeks. Tachyzoite antigens of the RH strain were detected 25 protein bands ranging 10 kDa-220 kDa of molecular weights with Coomassie blue stain. Toxoplcsma major antigens corresponding to n of 24 kDa, 27 kDa,30 kDa, 35 kDa, 38 kDa were recognized by IgG and IgM antibodies. Excretory-secretory antigens present in culture supernatant with M. W. of 20, 30 kDa and in infected mouse peritoneal fluid with M.W. of 33 (P30), 45 kDa. When RH tachyzoite antigen was probed with different mice sera immunized with 2 strains of T gondii, the IgG antibody bud of Fukaya and Beverly strain (8 week-serum) is identical to those of RH strain. It is considered that the 30 kDa polypeptide detected in excretory- secretory materials and Iysate was important major antigen of T gondii (RH).

• Parasites, Hosts and Diseases
• /
• v.35 no.4
• /
• pp.251-258
• /
• 1997

Tachyzoite antigens of Toxoplosnc gondii (RH) were partially purified by immunoaffinity chromatography. The cultivated ToxopLusmc in uiuo (mouse) and in nitro (Hep-2 cell) and peritoneal fluid of T. Bondii infected mice were collected for antigen analy- sis. Tachyzoite antigens collected from infected mouse showed positive bands of 76 kDa, 70 kDa,64 kDa, 53 kDa, 46 kDa, 44 kDa, 41 kDa, 35 kDa, 25 kDa, 18 kDa, and 13 kDa on immunoblot with anti-Toxoplcsmn rabbit sera, and those from infected Hep-2 cells revealed reactive bands of 70 kDa,64 kDa,53 kDa,35 kDa,28 kDa, and 13-10 kDa. After applying to an IgG-Sepharose column, two elusion peaks, E-1 and I-2 fractions, were obtained from both soluble antigen of T. gondii and the peritoneal fluid of infected mice, respectively. Immunoblots of soluble antigen with immunized rabbit sera revealed positive bands of 97 kDa, 63 kDa, 53 kDa and 35 kDa from I-1 fraction and 53 kDa and 35 kDa from I-2. In the case of the eluted peaks from mice peritoneal fluid, E-1 showed protein bands of 84 kDa,76 kDa,53 kDa and 29 kDa bands and 53 kDa and 45 kDa from I-2 on immunoblots. Serum IgG antibody titer of mice immunized with T gonnii tachyzoites was increased on 1 week after booster immunization when analysed by ELISA using crude antigen, while it was elevated on 3 weeks after booster immunization by ELISA using puri- fied antigen.

• Korean Journal of Microbiology
• /
• v.31 no.3
• /
• pp.230-236
• /
• 1993

The isocitrate lyase extracted from Micrococcus luteus was purified 38.8 folds with the overall yield of 10.2%, by the ammonium sulfate fractionation, DEAE-cellulose, 1st Sephadex G-200 and 2nd Sephadex G-200 column chromatography. The purified enzyme showed to be a single protein band by polyacrylamide gel electrophoresis. The molecular weight of the purified enzyme was estimated 60,000 by the SDS-polyacry]amide gel electrophoresis. The apparent Michaelis constant, Km value for isocitrate was 0.95 mM. The optimum pH and temperature of the purified enzyme were pH 7.5 and $40^{\circ}C$, respectively. The enzyme was activated by $Mg^{2+}$ and inhibited by $Mn^{2+}$, $Ca^{2+}$, $Cu^{2+}$, $Zn^{2+}$ and $CO^{2+}$. In addition, the activity of isocitrate lyase was increased by glutathione and 2-mercaptocthanol at 5 mM and cysteine at I mM.

• Journal of the Korean Chemical Society
• /
• v.32 no.4
• /
• pp.377-384
• /
• 1988

Carotenoprotein from Salmo Salar eggs was purified and characterized by CM-cellulose, 50% $(NH_4)_2SO_4$, DEAE-cellulose and sephadex G-75 column. The chromoprotein had a spectrum with ${\lambda}_{max}$ 409, 540 and 580nm in p-buffer (pH 7.0) at initial step. Molecular weights by sephadex G-200 gel filtration were 50, 200 and 26,000 daltons. SDS-PAGE analysis showed a structure with four identical subunits (12,500 daltons). Its sample retained a small amount of carbohydrates and lipids. Amino acids were analyzed, and mannose, galactose and glucosamine also were identified. Carotenoid extacted with acetone was found to be astaxanthin ester by partition test, epoxy test, iodine test, allylic test, reduction, acetylation, uv/vis, ir and nmr datas. Stearate (47.9%) and palmitate (21.4%) were predominant fatty acids in the astaxanthin ester.

• Parasites, Hosts and Diseases
• /
• v.27 no.1
• /
• pp.15-22
• /
• 1989

To observe antibody changes after praziquantel treatment in paragonimiasis, a total of 46 serum samples from 13 serologically diagnosed patients was collected for 4~28 months. The specific antibody (IgG) levels were measured by enzyme- linked immunosorbent assay (ELISA). All but one patient who needed retreatment became symptom-free within a week. Antibody levels were dropped near to or below a cut-off absorbance (abs.) of 0.25 in varying intervals from 4 to 18 months. Of 9 patients who were retested within 3 months, 5 revealed temporary elevation of antibody level. After the elevation, the levels be낙an to decline slowly to negative ranges. If treated earlier after symptoms developed, the temporary elevation did not occur and intervals to negative conversion were shorter. By sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) /immunoblot, antigen-antibody reactions in individual patient faded gradually without significant changes in reacting antigen bands.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.21 no.6
• /
• pp.725-730
• /
• 1992

The internal invertase was purified from cell free extract of Rhodosporidium toruloides IFO 0559-M-919 by acid precipitation, ion-exchange chromatography and gel filtration to the unique enzyme protein on disc electrophoresis. We have found out that molecular weight of purified internal invertase was 90,000 by gel filtration and the purified enzyme was protein with 4 homogeneous subunits appearing as single band of 22,000daltons on SDS-polyacrylamide gel electrophoresis.

• Biomedical Science Letters
• /
• v.2 no.1
• /
• pp.1-12
• /
• 1996

Antigenic components reacting with IgE and IgG antibodies were localized in muscular layer of adult and of larva, sparganum. But the antigenic components inducing IgG were localized at tegument and parenchyma in addition to muscular layer in adult and sparganum. Also in sparganum, the surface of calcareous corpuscles of parenchyma showed immunoreactivity to IgG antibody. However antigenic components inducing IgE antibody were not localized in tegument and parenchyma, but in adult worm, we observed the immunopositive reaction at the lining of vitelline follicles in mature proglottis and on surface of egg shell within uterus of graved proglottis. By the method of immunogold-labelling, we observed the location of antigenic particles in tegument of sparganum. The density of antigenic particles inducing IgG was higher than that of antigen particles inducing IgE in syncytial tegument, tegument cells. A total of 43 and 36 protein bands were resolved from crude extracts of adult and sparganum, respectively, by SDS-PAGE. 34 bands from crude extracts of adult and larva were migrated to same positions. By EITB, 21 bands of 44 bands in adult were recognized with IgG antibody, and also 21 bands of 36 bands in sparganum. 13 bands of them were common antigenic components both in the adult worm and sparganum. Because 19 bands of 44 bands in adult worm were reacted with IgE antibody, they were IgE antigenic component. In sparganum, 13 bands were IgE antigenic components. 9 bands of them were common antigenic component inducing IgE antibody in both a-dult and sparganum. 3 bands of antigenic component recognized by IgE and IgG antibody were nonspecific antigen in both adult and sparganum of Spirometra erinacei.

• 한국생물공학회:학술대회논문집
• /
• 2003.10a
• /
• pp.499-503
• /
• 2003

In the study, we have obtained modified cellulase with higher cellulose degradation activity by molecular evolution method. Cellobiohydrolase(CBH I ) gene of Trichorderma reerri has been used. Cellulase mutant 228-G2 was selected and the activity of cellulase mutant 228-G2 was increased by 300% compared to original CBH I The 17 among 1542bases were found to be modified with mutant 228-G2.

• Korean Journal of Animal Reproduction
• /
• v.20 no.2
• /
• pp.215-221
• /
• 1996

This study was carried out to develop an immunoassay for the diagnosis of the pregnancy and ovarian function of domestic animals. Using 2E92C10 monoclonal antibody(McAb) generated against estrone-3-glucuronide(E1-3-G) and appeared a high cross-reactivity with estrone-3-sulfate(E1-3-S), chemiluminesence immunoassay (CIA) to detect E1-3-S was developed. 2E92C10 McAb cross-reacted with E1-3-S (30%) was purified from ascites fluid using protein G sepharose gel column. The purity of purified antibody fraction was monitored by SDS-PAGE and was better compared to that of crude ascite fluid. The soild and liquid phase CIA for E1-3-S were established utilizing 2E92C10 antibody and E1-3-G-ABEI conjugate used as a tracer. As the results, the titer of 2E92C10 antibody was 5g/ml in soild phase and 1:2000 in liquid phase. The sensitivity on soild and solid phase CIA were about 200 pg/ml. These results indicate that CIA for measurement of E1-3-S was successfully developed by using ant-E1-3-G McAb cross-reacted with E1-3-S and could be usefully used to research this area.

• Korean Journal of Food Science and Technology
• /
• v.31 no.4
• /
• pp.899-905
• /
• 1999

The lectins from mucilaginous jelly and green epidermis of Aloe vera were isolated by gel and affinity chromatography. The molecular weights of the lectins were determined by SDS-PAGE. The molecular weights of the lectins from mucilaginous jelly isolated by Sephadex G-100 were 58.7 kD and 33.3 kD, and that isolated by acid-treated Sepharose 4B was 176.4 kD. The molecular weights of the lectins from epidermis isolated by Sephadex G-100 were 221.1, 54.0 and 32.5 kD respectively. And that isolated by acid-treated Sepharose 4B was 222.0 and 158.0 kD. The agglutinating activity of lectin from jelly was inhibited by D-galactose, lactose and D-galactosamine, but that from epidermis was not inhibited by lactose. The activity was stable at the pH range of $7.0{\sim}9.0$ and at the temperature $0{\sim}60^{\circ}C$.