• Title/Summary/Keyword: antibody engineering

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Molecular Screening and Characterization of Antiviral Potatoes

  • Tripathi, Giriraj;Li, Hongxain;Park, Jae-Kyun;Park, Yoon-Kyung;Cheong, Hyeon-Sook
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.89-95
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    • 2006
  • Potato plants carrying the Ry gene are extremely resistance to a number of potyviruses, but it is not known which variety expressed the resistance. In this investigation, combined classical and molecular techniques were used to identify virus resistance potatoes. Mechanical inoculation of 32 varieties of Korean potato cultivars, with potato virus Y (PVY), induced various symptoms, such as mosaic, yellowing, necrosis, mottle, vein clearing and vein bending. Different virus spreading patterns were observed, such as highly sensitive, moderate and resistant to $PVY^o$ inoculated leaves in different cultivars. From the results of double antibody sandwich-enzyme links immunosorbant assays (DAS-ELISA), coupled with reverse transcription polymerase chain reaction (RT-PCR), Winter valley and Golden valley were found to be highly susceptible and resistant cultivars to $PVY^o$ respectively. TEM was used as a complementary method to conform the localization of the virus in leaf tissues. TEM detect virus particles in Golden valley, where, ELISA and RT-PCR were unable to detect the CP gene. However, the interior part of the tissues was severely deformed in $PVY^o$ infected Winter valley, than Golden valley The Ry gene is involved in an induced response in $PVY^o$ infected Golden valley plants. The methods described in this study could be applied for the screening and development of antiviral potatoes.

Cloning and Expression of hpaA Gene of Korean Strain Helicobacter pylori K51 in Oral Vaccine Delivery Vehicle Lactococcus lactis subsp. lactis MG1363

  • Kim Su-Jung;Jun Do-Youn;Yang Chae-Ha;Kim Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.318-324
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    • 2006
  • In order to develop an oral vaccine to prevent H. pylori infection, we have expressed the hpaA gene of H. pylori K51 isolated from Korean patients, encoding 29-kDa HpaA that is known to be localized on the cell surface and flagella sheath, in a live delivery vector system, Lactococcus lactis. The hpaA gene, amplified by PCR using the genomic DNA of H. pylori K51, was cloned in the pGEX-2T vector, and the DNA sequence analysis revealed that the hpaA gene of H. pylori K51 had 99.7% and 94.8% identity with individual hpaA genes of the H. pylori 26695 strain (U.K) and the J99 strain (U.S.A). A polyclonal anti-HpaA antibody was raised in rats using GST-HpaA fusion protein as the antigen. The hpaA gene was inserted in an E. coli-L. lactis-shuttle vector (pMG36e) to express in L. lactis. Western blot analysis showed that the expression level of HpaA in the L. lactis transformant remained constant from the exponential phase to the stationary phase, without extracelluar secretion. These results indicate that the HpaA of H. pylori K51 was successfully expressed in L. lactis, and suggest that the recombinant L. lactis expressing HpaA may be applicable as an oral vaccine to induce a protective immune response against H. pylori.

The Development of Blood Bank Management Program (혈액 은행 전산 처리 프로그램의 개발)

  • Kim, Jong-Won;Lee, Seung-Kuk;Han, Kyou-Sup;Kim, Jin-Q;Cho, Han-Ik;Kim, Sang-In
    • Proceedings of the KOSOMBE Conference
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    • v.1989 no.05
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    • pp.75-76
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    • 1989
  • The blood bank is a field of clinical pathology which requires the most accuratemaintenanceofrecording. Because the mistake in it is directly related to a patient's life. So, the computerization of the blood bank is urgent to maintain a log blook arid to compare the patient's current data with past result. We developed the blood bank management program using 32 bit minicomputer. This is composed of 4 parts; a management of routine test result, special test result, the blood issue and statistics. The management of routine test result handles the patient's information and blood typing and compares above results with the past one of same patient. The management of special test result are for special immunohematologic tests like an irregular antibody, Coombs' test, and etc. Blood issue part records the type of the blood bag, component, and the name of issuer. Statistic part are made to get statistics of each day and each month by the blood type, and the type the blood component. The program is secured by the maintenance of operator's operation history and thu provision of the security code to each operator, without which no one can enter the system and after the content. So the stability and reliability of the data is obtained. This program will be upgraded for bar-code using system in the near future.

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Serologic survey of the ruminant bacterial infectious diseases in farmed deer and wild water deer in Jeonbuk province (사육사슴 및 야생고라니에서 소 세균성 전염병에 대한 혈청학적 연구)

  • Jo, Young-Suk;Chung, Yun-Shin;So, Seung-Young;Seol, Min-Suk;Cho, Ho-Seong;Kim, Bum-Seok;Lim, Chae-Woong
    • Korean Journal of Veterinary Service
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    • v.33 no.3
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    • pp.249-254
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    • 2010
  • Deer can be one of the susceptible animals to bovine infectious diseases, and thus, may play a role either as a reservoir or amplifier host for spreading the diseases to other species such as cattle and goat. This study was conducted to determine the serum antibodies to bacterial infectious diseases for brucellosis, tuberculosis (TB), paratuberculosis (Johne's disease) in deer. Serum samples were randomly collected from 78 deer from 31 farms at Jeonbuk province, and 7 wild water deer from Jeonbuk wild animal treatment center during 2005 to 2007, respectively. Four farm deer (5.1%) showed antibodies to tuberculosis using Antigen Rapid Bovine TB Ab Test Kit. One elk (1.3%) and one wild water deer had antibodies for paratuberculosis. Antibody against Brucellosis was not detected in tube agglutination test (TAT) and enzyme-linked immunosorbent assay (ELISA). These data suggest that caution should be applied to inspection of velvet, deer blood and meat for human consumption from deer because of zoonotic bacterial diseases in deer. In addition, farmed deer can be a transmissible host for zoonotic disease to diary or raising farm.

A Study on the production of Monoclonal Antibodies against Rhodotrodula rubra (효모양진균의 단크론 생성)

  • Ryeom K.;Kim S. C.;Lee J. H.
    • Journal of environmental and Sanitary engineering
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    • v.4 no.1 s.6
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    • pp.17-28
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    • 1989
  • Having studied the production of monoclonal antibodies for developing a diagnosis medicine which shall be detected by a high-sensitivity test by using Rhodotorula rubra as a fungi-host which had been extracted through biochemical tests and follow-up examinations on Yeast-like fungi obtained from pulmonary tissues of pulmonary tuberculosis patients who had been in Kong ju National Tuberculosis Hospital from Jun. to Dec. in 1987, I. have gained such results as follows: 1. The fusion rate was influenced by feeder cell layers, cell density and time required to the cell fusion with cells in myelona subculture. 2. The fusion rate did not show any significant difference when the cell was applyed with two molecular weights, i.e., 1500 and 4000, of polyethylene glycol. 3. Fused cells after the addition of HAT selection media were bright and round, whereas unfused myelona cells and spleen cells were shrunk and granulated. 4. The cell fusion rate turned out to be about $57.2\%$(150 wells / 264 wells). 5. $10\%$(15 wells / 150 wells) of the positive reaction was detected in monoclonal antibody screening. 6. The titer which had reacted positively to Rhodotorula rubra fungal-host was 800 times in density after the gradual dilution of the produced monoclonal antibodies with Indirect ELISA method. 7. The Strongest specific reaction came out after the peroxidase labelled anti-human Immunogobulin had been applyed to Rhodotorula rubra for activating its nature after making drift with Carbonate-bicarbonate buffer (pH 9.6) and drying completely.

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Molecular Cloning of the Gene for $\alpha$-Acylamino-$\beta$-lactam Acylhydrolase from Acetobacter turbidans by Immunochemical Detection Method (면역화학적 방법에 의한 Acetobacter turbidans의 $\alpha$-Acylamino-$\beta$-lactam Acylhydrolase의 유전자 클론화)

  • Nam, Doo-Hyun;Dewey D.Y. Ryu
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.363-368
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    • 1988
  • Molecular cloning of gene for $\alpha$-acylamino-$\beta$-lactam acylhydrolase (ALAH) III from Acetobacter turbidans has been attempted by immunochemical detection method, in which polyclonal antibody from mouse Balb/c against this enzyme was employed as a probe. As a cloning vector, λ gtll was chosen for this purpose. Two positive clones has been selected from genomic libraries of A. turbidans, which had somewhat different binding affinities on anti-ALAH III umm and anti-$\beta$-galactosidase. By restriction analysis, both clones has been turned out to lose one of EeoRI sites. From these results, it concluded that deletion of DNA between lacZ gene and inserted DNA has occurred during replication of these clones in host cells.

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Myelination by co-culture of neurons and schwann cells and demyelination by virus infection (뉴런세포와 슈반세포의 공동배양에 의한 수초화와 바이러스 감염에 의한 탈수초화)

  • Sa, Young-Hee;Kweon, Tae Dong;Kim, Ji-Young;Kim, Hyun Joo;Lee, Bae Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2018.10a
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    • pp.448-451
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    • 2018
  • The purpose of this study was to investigate the developmental process of myelination by neuron and Schwann cell cultures and the development of demyelination by herpes simplex virus-1 infection by electron microscopy and molecular biological analysis. The dorsal root ganglion (DRG) was isolated from the mouse embryo and Schwann cells and neuronal cells were cultured in vitro. Neuronal cells treated with mitotic inhibitors and purified Schwann cells were co-cultured together to induce myelination. The herpes simplex virus-1 was infected with the co-cultured cells, and the demyelination was induced. The myelin protein zero (MPZ) antibody, which means the presence of myelin formation, was used and electron microscopy was used to observe the development of myelin and dehydration.

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Effect of Kp, an Antitumor Protein-Polysaccharide from Mycelial Culture of Phellinus Linteus on the Humoral lammune Response of Tumor-Bearing ICR Mice to Sheep Red Blood Cells

  • Chung, Kyeong-Soo;Kim, Shin-Sook;Kim, Hee-Soo;Kim, Kye-Yang;Han, Man-Woo;Kim, Kil-Hyun
    • Archives of Pharmacal Research
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    • v.16 no.4
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    • pp.336-338
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    • 1993
  • The immunomodulating activity of Kp, an antitumor protein-polysacchanide preparation from the shake-cultured mycelia of Phellinus linteus, was investigated in ICR mice subcutaneously implanted wit $1\times10^6$ cells of sarcoma 180. The mice were intraperitoneally administered with Kp at a does of 100 mg/kg once daily for five consecutive days starting from 24 hrs after the tumor implantation. Ten days after the last injection, the mice were immunized with $1\times10^7$ or $4\times10^8$ sheep red blood cells (SRBC) and five days later, the antibody-forming immune response were assessed by direct hemolytic plaque assay. To an immunization does of $1\times10^7$ SRBC, the Kp-treated mice elicied a successful humoral immune response despite the turmor-burden and produced $259\times10^3$ plaque-forming cells (PFC)/spleen, while the corresponding tumor-bearing control mice showed virtually no reponse $(2.0\times10^3$ PFC/spleen) (the stimulation index=129.5). However, to an immunization dose of $4\times10^8$ SRBC, both of the control mice and Kp-treated mice showed almost the same level of strong humoral immune response. From these data it is clear that Kp effectively restores the humoral immune response of the turmor-bearing ICR mice.

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Purification and Characterization of NADPH-Dependent Cr(VI) Reductase from Escherichia coli ATCC 33456

  • Bae, Woo-Chul;Lee, Han-Ki;Choe, Young-Chool;Jahng, Deok-Jin;Lee, Sang-Hee;Kim, Sang-Jin;Lee, Jung-Hyun;Jeong, Byeong-Chul
    • Journal of Microbiology
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    • v.43 no.1
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    • pp.21-27
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    • 2005
  • A soluble Cr(VI) reductase was purified from the cytoplasm of Escherichia coli ATCC 33456. The molecular mass was estimated to be 84 and 42 kDa by gel filtration and SDS-polyacrylamide gel electrophoresis, respectively, indicating a dimeric structure. The pI was 4.66, and optimal enzyme activity was obtained at pH 6.5 and $37^{\circ}C$. The most stable condition existed at pH 7.0. The purified enzyme used both NADPH and NADH as electron donors for Cr(VI) reduction, while NADPH was the better, conferring 61% higher activity than NADH. The $K_m$ values for NADPH and NADH were determined to be 47.5 and 17.2 umol, and the $V_max$ values 322.2 and 130.7 umol Cr(VI) $min^{-1}mg^{-1}$ protein, respectively. The activity was strongly inhibited by N-ethylmalemide, $Ag^{2+},\;Cd^{2+},\;Hg^{2+}$, and $Zn^{2+}$. The antibody against the enzyme showed no immunological cross reaction with those of other Cr(VI) reducing strains.

Effects of dietary supplementation with different levels and molecular weights of fungal β-glucan on performances, health and meat quality in broilers

  • Kovitvadhi, Attawit;Chundang, Pipatpong;Tirawattanawanich, Chanin;Prathumpai, Wai;Methacanon, Pawadee;Chokpipatpol, Krith
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.10
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    • pp.1548-1557
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    • 2019
  • Objective: To investigate the effects of dietary supplementation with different levels and molecular weights of fungal ${\beta}$-glucan on productive performances, health, carcass traits and meat quality in broilers. Methods: Two hundred and ten of one-day-old chicks with equal sex were assigned to seven experimental groups in $2{\times}4$ factorial arrangement. These groups were supplemented with (0, 10, 30, and 60 ppm) of molecular weight 1-3, 1-6 ${\beta}$-glucan (low or high). High molecular weight ${\beta}$-glucan (H: 943 kDa) was obtained from Ophiocordyceps dipterigena BCC 2073, whereas H with ${\gamma}$-Irradiation treatment was performed to achieve low molecular weight ${\beta}$-glucan (L: 8 kDa). Results: There was no statistical significance in productive performances, apparent digestibility and interaction between fixed factors along 42 days of experiment (p>0.05). A higher caecal amylase activity was present in the group that received L, while there was a dramatic decrease in H and the control groups, respectively (p<0.05). The increase of supplemental dose increased caecal amylase activity (p<0.05). Immunomodulatory effects from L was revealed by the marked increase of phagocytic activity, relative weight of thymus and bursa of fabricius (p<0.05). Similarly, the additive dose at 30 ppm provided the same results, whereas the only significant difference with supplementation at 60 ppm was an increase in phagocytic activity (p<0.05). Interestingly, villi height of broilers fed L was higher than other groups (p<0.05). The treatments did not influence haematology, blood chemistry, antibody production level against vaccination, carcass traits and meat quality (p>0.05). Conclusion: The supplementation of L at 30 ppm was suggested to achieve benefits of immune modulation without adverse effects on other parameters.