• Journal of Life Science
• /
• v.20 no.9
• /
• pp.1294-1298
• /
• 2010

White spot syndrome virus (WSSV) is one of the most virulent viral agents threatening the penaeid shrimp culture industry. This study was carried out to evaluate the susceptibility of the sand shrimp, Crangon affinis, to WSSV as an alternative experimental model. WSSV caused 100% mortality in C. affinis within 7 days after experimental infection by immersion. Based on challenge studies, it was confirmed that C. affinis could be a potential host in WSSV transmission. Also, the neutralization of WSSV was carried out using an antiserum raised against recombinant envelop protein rVP466 to evaluate the WSSV infection mechanism. A constant amount of WSSV (at $1{\times}10^4$ diluted stocks) was incubated with various amounts of antiserum and then mixed to 20 l reservoir for the immersion challenge of C. affinis for neutralization. At 5 days post challenge, the shrimp in the positive control immersed in the immersion reservoir containing WSSV stock showed 100% mortality. The shrimps challenged with the 3 different mixtures of WSSV and rVP466 antiserum (1:0.1, 1:0.5 and 1:1) showed 100%, 68.8% and 68.8% mortality at 14 days post challenge, respectively. These results indicated that the antiserum raised against rVP466 could block WSSV infection in C. affinis. Therefore, this study confirmed that C. affinis can be naturally infected by WSSV as another potential host and that C. affinis can be used as an alternative experimental animal instead of penaeid shrimps.

• Korean Journal of Veterinary Research
• /
• v.16 no.1
• /
• pp.7-10
• /
• 1976

An agglutinability between naturally infected positive chicken serum of pullorum disease and hyperimmumized rabbit antiserum was compared. And the following results were obtained and summarized. 1. On the agglutinability, Salmonella pullorum antigen which irradiated gamma-ray was more excellent than another both formalized and heated antigen. 2. Time of judgemented as positive titer in the tube agglutination test to the naturally infected positive chicken serum was it most suitable for 12 hours at $37^{\circ}C$. 3. Agglutination titer of positive immune chicken serum against gamma-ray irradiate Salmonella pullorum were as 320~640x.

• Journal of Plant Biotechnology
• /
• v.4 no.1
• /
• pp.39-44
• /
• 2002

The fast-migrating cationic peroxidase isozyme, named RC3, was purified from rice (Oryza sativa cv. Nak-Dong) callus. Purification of the enzyme was accomplished by ammonium sulfate fractionation, CM-cellulose ionexchange chromatography, and Sephacryl S-100 gel filtration. The molecular mass of the enzyme was about 34 KDa as determined by SDS-PACE and 38 KDa by Sephacryl-100 gel filtration. The pI value of the enzyme was 8.9. Antiserum against RC3 was raised in rabbits, and anti RC3 antiserum reacted with RC3 isozyme by Ouchterlony double immunodiffusion. The optimum pHs and Km values of the enzyme for various substrates were determined. Kinetic studies with various substrates showed that RC3 had very low Km value of 0.01 mM for ferulic acid and ascorbic acid. However, the enzyme did not use esculetin as a substrate.

• Korean Journal of Animal Reproduction
• /
• v.10 no.1
• /
• pp.36-41
• /
• 1986

These experiments were carried out to obtain basic information necessary for sexing embryos by chromosomal analysis. To observe metaphase chromosomes, all embryos developed to blastocysts were cultured in Ho, pp. & Pitts' medium containing 0.001% Colcemid under the gas phase of 5% CO2 in air at 37$^{\circ}C$ for 2 hours. The sex chromosome of mouse embryos shown normal development after culture in medium containing H-Y antiserum (10%, v/v) and complement (20%, v/v) also was confimed by chromosomal analysis. The results obtained in these experiments were summarized as follows: 1. Among 89 mouse blastocysts, the number of embryos identified to have XX and XY chromosome was 22(25%) and 25(28%), respectively and 42(47%) embryos were not identified. 2. Of total 40 mouse balstocysts cultured in medium containing H-Y antiserum and complement, 23(58%) embryos which were able to be discriminated their sex chromosomes were identified to be XX bearing embryos. 3. Sex chromosomes of a number of embryos subjected to chromosomal analysis were not identified. This result may be due to absence or poor quality of metaphase spreads.

• KSBB Journal
• /
• v.5 no.4
• /
• pp.315-321
• /
• 1990

Rabbit anti-bovine heart PDH antiserum was raised against El(a, b) isolated from PDC, and then applied to detect Ela and Elb. Appropriate amounis of El were fractionated by SDS-PAGE and electrophoretically transferred to nitrocellulose membrane. The Ela and Elb on the membrane were incubated with anti-El antiserum and identified by GAR-HRP system. It has been found that the immunodetection sensitivity of Ela and Elb were directly proportional to the amount of antigen and transfer time. The lengthy transfer times increased the immunodetection sensitivity of Ela and Elb. The maximal detection sensitivity of Western blotting of Ela and Elb was achieved at 3.5 V/cm for 16-hour transfer under these experimental conditions.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.29 no.2
• /
• pp.214-219
• /
• 2014

We prepared antibodies from insect glycosaminoglycans (GAGs) and assayed the titer. Nine polyclonal antibodies against insect GAGs were raised for development of an ELISA in biological fluids (mice serum). The 3th booster collection of antiserum of BALB/c mice as a primarily antibody was assayed for titer determination by ELISA method. In sandwich ELISA of GAGs derived from Isaria sinclairii or other insects, antiserum from insect GAGs gave satisfactory results for so potent antibody(100: 1~1000:1) raising (manufacturing) agent in range of 10 ng/ml.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.150.2-150
• /
• 2003

Carnation mottle carmovirus(CarMV) was isolated from Lilium spp. in Korea. This isolate, CarMV, was done bioassay, which plants were Dianthus caryophyllus, Gomphrena globosa, Chenopodium amaranticolor, Dianths chinensis. CarMV was propagated on the leaves of Chenopodium amaranticolor with the crude-sap inoculation method and purified by Mossops method(1976). We produced antiserum against CarMV and analyzed the antiserum specificity with ELISA, Gel diffusion method, and Rapid Immunofilter Paper Assay (RIPA). From these results of the assay, RIPA method was simple and rapid for CarMV detection. We have established successfully the CarMV detection system. CarMV coat protein gene was amplified by RT-PCR with specific primers and sequencing analysis was done.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.127.2-127
• /
• 2003

YMV-K strains were purified from D. oppostita Thunb. tv. Dung-Gun-Ma showing mosaic symptom on their leaves. YMV-K strains were filamentous particles of 780nm in length and induced cytoplasmic disorder such as inclusion body formation. Nucleotide and amino acid sequences of 5'-UTR, Pl and CP of YMV-K strains shared 80.8, 64.7 and 98.3％ identity respectively to JYMV J1 in the mean value. Purification of YMV-K strains according to JYMV purification method(S. Fuji) was conducted to product antiserum. With antiserum against YMV-K strains, the Various diagnosis methods such as IC-RT-PCR, DIBA, RIPA and indirect-ELISA were used to detect YMV-K strains in Chinese yam plant.

• Proceedings of the Korean Society of Life Science Conference
• /
• 2001.11a
• /
• pp.34-34
• /
• 2001

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.27 no.6
• /
• pp.1152-1159
• /
• 1998

In order to develop more rapid and reproducible analysis of biotin known as vitamin H, attempts were made to establish the condition for enzyme linked immunosorbent assay(ELISA) compared with traditional microbiological assay(MBA). Antibiotin and antiserum were obtained from the immunized rabbits injected with emulsion of biotin KLH conjugate and Freund's adjuvant. The antiserum showed cross reactivity on biocytin, a derivative of biotin, which is converted to biotin in intestine, at the rate of 177%(median inhibitory concentration(IC50)=12.58ppb), but not on other derivatives such as desthiobiotin, diaminobiotin and 2 imino biotin. Specific antibody for biotin was purified from the antiserum through protein A column and desalting column. The conditions of competitive direct ELISA (cdELISA) were established. Detection range of biotin concentration by cdELISA was 0.01∼300ng/ ml(ppb). In the spike test with milk, fruit flake and pine carrot juice, the correlation coefficient between two methods of MBA and ELISA was reliably consistent at the value of r=0.992. But detection of biotin by microbiological assay(MBA) was rather restricted in range and nonspecific. Detection range of biotin by MBA was 0.1∼0.5ng/ml(ppb). It showed cross reactivities on biocytin and desthiobiotin at the rate of 80.1% and 66.7%, respectively. In conclusion, ELISA revealed a significant improvement compared with MBA for the biotin detection in terms of sensitivity, detection range and cross reactivity. In addition, a variety of samples could be analyzed rapidly and conveniently at one time by using ELISA. These results strongly suggest that the ELISA is very promising for the practical application to detect biotin contents in a wide range of food stuffs.