• Title/Summary/Keyword: Antibody Engineering

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Enzyme-linked Immunosorbent Assay Strip Sensor for Rapid Detection of Staphylococcus aureus (Staphylococcus aureus 신속 검출을 위한 효소면역측정 스트립 센서)

  • Park, So Jung;Kim, Young-Kee
    • Applied Chemistry for Engineering
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    • v.22 no.5
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    • pp.522-525
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    • 2011
  • In this study, an established enzyme-linked immunosorbent assay and immuno-chromatography technique are combined to fabricate an immuno-strip sensor for the detection of S. aureus. The immuno-strip is manufactured by using four different functional membranes. The capture antibody is immobilized on the nitrocellulose membrane due to the high affinity and the capillary action through porous membranes induces a flow of sample. A colorimetric signal is appeared according to the enzyme reaction and is analyzed by the digital camera (qualitative analysis) and home-made image analysis software (quantitative analysis). Under the optimal conditions, samples with S. aureus in the range of $2.7{\times}10^4{\sim}2.7{\times}10^7CFU/mL$ can be detected by the colorimetric method within 30 min.

Expression of Immunologically Active Porcine Recombinant TGF-${\beta}1$ Precursor Protein in Baculovirus System

  • Lim, Hyun;Kim, Pyeung-Hyeun;Chun, Gie-Taek;Choi, Eui-Yul;Yie, Se-Won
    • Journal of Microbiology
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    • v.35 no.4
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    • pp.341-346
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    • 1997
  • In order to express recombinant porcine TGF-${\beta}1$ protein in a baculovirus expression system the entire TGF-${\beta}1$gene containing extra amino acids at the N terminus was cloned into pFBa and pFBb of the Bac-To-$Bac^{TM}$ baculovirus expression system. One of the clones contained 106 extra amino acids and was designated pFBa-106 TGF-${\beta}1$, and the other had 28 extra amino acids and was designated pFBb-28 TGF-${\beta}1$. The orientation of the gene was identified with restriction enzyme mapping and PCR with internal TGF-${\beta}1$ primers. Sf-9 cells were infected at a m.o.i. of 10 by the recombinant viruses generated from the two expected sizes of 55 kD and 46.4kD. these precursor forms of TGF-${\beta}1$ with a polyclonal antibody against human TGF-${\beta}1$. No mature form of TGF-${\beta}1$ protein was detected on SDS gels and an immunoblot indicated that TGF-${\beta}1$ precursor is not properlu processed in insect cells.

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Effect of Dehydration and Rehydration of the pH-Sensitive Liposomes Containing Chimeric gag-V3 Virus Like Particle on Their Long-term Stability

  • Chang, Jin-Soo;Park, Myeong-Jun;Kim, Tae-Yeon;Woo, Gyu-Jin;Chung, Soo-il;Cheong, Hong-Seok
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.66-71
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    • 1999
  • One of the practical limitations with the use of liposomes for delivery of the pharmaceutical substances such antigens is that liposomes are relatively unstable in storage. In order to extend the stability of liposome in storage without affecting their functional activity, solution-type liposomes were dehydrated to form a structurally intact dry liposomes. Comparative immunological evaluation was carried out for both dry and solution-type liposomes containing gag-V3 chimera, consequently it was found that dry liposomes elicited both humoral and cellular response as efficiently as solution-type liposemes did against the same gag-V3 antigen. Especially, long-term stability of the liposomes was remarkably enhanced by the dehydration made to loposomes without a significant change in its ability to elicit immune response in vivo. These results indicate that dry pH-sensitive liposome may become an effective delivery and adjuvant system for general vaccine development.

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Clearance of False-positive Antigen-Antibody Reactions of a Diagnostic Antigen Production in Escherichia coli with Human Sera

  • Noh, Kap-Soo;Kim, Jong-Wan;Ha, Suk-Hoon;Yoo, Wang-Don;Jeon, Weong-Joong;Kim, Hyun-Su
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.63-65
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    • 1999
  • Although many pharmaceutically useful proteins are produced in E. coli expression system, it is very are rare for the system to be used in the production of diagnostic antigen due to a major problem, i.e., false-positive reaction of e. coli host-derived proteins contaminating purified diagnostic antigen with human sera. The N (nucleocapsid) protein of Seoul virus causing haemorrhagic fever with renal syndrome (HFRS) was produced in E. coli BL21 (DE3), and used for the detection of N protein-specific antibodies in human sera. Using the N protein as a diagnostic antigen of HFRS, the false-positive reaction was cleared by merely mixing the test sera with the extract of E. coli host strain not harboring expression plasmid.

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Study on Change of Poly ADP Ribose Polymerase in the Rat with Thrombotic Stroke by Full Wave Cockroft Walton method's Transcranial Magnetic Stimulation

  • Kim, Whi-Young;Kim, Jun-Hyoung
    • Journal of Magnetics
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    • v.19 no.1
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    • pp.20-27
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    • 2014
  • This study examined the relationships between protein expression and Poly ADP ribose polymerase in brain cell death in brains damaged by thrombotic stroke and treated with the Full Wave- Cockroft Walton (FWCW) method of Transcranial Magnetic Stimulation (TMS). The two-way switching element for TMS drove a half-bridge inverter of the current resonance of direct current voltage (+) and direct current voltage (-), and the experiment was conducted by stimulating the mice with thrombotic stroke through a range of pulses. Thrombotic stroke was caused of ligation of the common carotid artery of male SD mice, and blood reperfusion was conducted five minutes later. Protein expression was examined in immune reaction cells, which reacted to an antibody to Poly ADP ribose polymerase in the cerebrum cells, and western blotting. Observations of the PARP changes after thrombotic stroke showed that the number of Poly ADP ribose polymerase reactions were significantly lower (p < 0.05) in the group treated with TMS of the FWCW than the group with thrombotic stroke 24 hours after its onset. The application of FWCW-TMS helped prevent the necrosis of nerve cells and might prevent the brain damage that occurs as a result of thrombotic stroke, and improve the function recovery and disorder of brain cells.

A Study on the Antigen Characteristics of Rhodotorula rubra (Rhodotorula rubra의 항원특성에 관한 연구)

  • Kwon, Hyuk-Ku;Lee, Jang-Hoon;Ryeon, Kon
    • Journal of Environmental Health Sciences
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    • v.28 no.5
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    • pp.28-34
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    • 2002
  • Antigenicity of Rhodotrula rubra isolated from pulmonary tissue of pulmonary tuberculosis patients was studied by means of agglutination reaction with R. rubra whole cell antiserum. And the serological reactivity of crude polyfac charide from R. frubra, Candida albicans, Candida tropicalis, Candida, glabrata, and Saccharomyces cerevisiae ATCC 26603 with antiserum to R. rubra whole cell was studied by means of immunodiffusion test. R. rubra showed stationary phase after 48h when it was cultured in GYEP broth. While agglutinogen titer was 1:64 at lag phase, agglutinogen titer was 1 :256 after 20h. After growth of R. rubra on different 11 media, nutritional environment showed similar agglu-tination reartivity. The agglutinogen titer of C. albicans, C. tropicalis, C. giabrata, which were isolated from patient's expectoration, to R. rubra antiserum by means of agglutination reaction were 1:16, respectively. But, Sacch. cervisiae ATCC26603 was negative. Those results were lower than that of R. rubra agglutinogen titer 1:256. As a result of immu-nodiffusion test with crude polysaccharide extracted from cell wall of R. rubra, C. albicans, C. tropicalis, C. glabrata, Sacch. cervisiae ATCC26603, precipitin line was found only with R. rubra, of which antibody titer was 8.

Production of Monoclonal Antibody about Specific Key Enzyme of Hyoscyamine $6{\beta}-Hydroxylase$ (H6H) in Scopolia parviflora

  • Kang, Young-Min;Jung, Hee-Young;Kang, Seung-Mi;Jin, Byung-Rae;Lee, Sang-Chul;Lee, Byung-Hyun;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.2
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    • pp.135-140
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    • 2004
  • Total RNAs were isolated from cultured roots of Scopolia parviflora, $poly(A)^+$ RNA was obtained through the mRNA purification, cDNA library of Hnh6h was constructed. Recombinant baculoviruses in Spodoptera frugiperda (Sf) cells were constructed by use of the transfer vector pBacPAK, which has the AcNPV sequence under the polyhedrin promoter. The expression vector carrying Hnh6h gene was transferred to S. parviflora and obtained transgenic hairy root lines. Our results confirmed the over expression of the H6H protein was used by anti-pBacPAK about cDNAs of S. parviflora. This study will served for production of tropane alkaloids by metabolic engineering.

Evidence for the Ras-Independent Signaling Pathway Regulating Insulin-Induced DNA Synthesis

  • Jhun, Byung-H.
    • BMB Reports
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    • v.32 no.2
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    • pp.196-202
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    • 1999
  • The existence of the Ras-independent signal transduction pathway of insulin leading to DNA synthesis was investigated in Rat-1 fibroblasts overexpressing human insulin receptor (HIRc-B) using the single-cell microinjection technique. Microinjection of a dominant-negative mutant $Ras^{N17}$ protein into quiescent HIRc-B cells inhibited the DNA synthesis stimulated by insulin. Microinjection of oncogenic H-$Ras^{V12}$ protein ($H-Ras^{V12}$) (0.1 mg/ml) induced DNA synthesis by 35%, whereas that of control-injected IgG was induced by 20%. When the marginal amount of oncogenic H-$Ras^{V12}$ protein was coinjected with a dominant-negative mutant of the H-Ras protein ($Ras^{N17}$), DNA synthesis was 35% and 74% in the absence and presence of insulin, respectively. This full recovery of DNA synthesis by insulin suggests the existence of the Ras-independent pathway. The same recovery was observed in the cells coinjected with either H-$Ras^{V12}$ plus H-$Ras^{N17}$ plus SH2 domain of the p85 subunit of PI3-kinase ($p85^{SH2-N}$) or H-$Ras^{V12}$ plus H-$Ras^{N17}$ plus interfering anti-Shc antibody. When co-injected with a dominant-negative H-$Ras^{N17}$, the DNA synthesis induced by the Ras-independent pathway was blocked. These results indicate that the Ras-independent pathway of insulin leading to DNA synthesis exists, bypassing the p85 of PI3-kinase and Shc protein, and requires Rac1 protein.

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Rapid Detection Kit for Salmonella typhimurium (살모넬라 식중독균 신속 검출용 간이 진단키트)

  • Kim, Gi-Young;Yang, Gil-Mo;Park, Saet-Byeol;Kim, Yung-Hwun;Lee, Kang-Jin;Son, Jae-Yong;Kim, Hyuck-Joo;Lee, Sae-Rom
    • Journal of Biosystems Engineering
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    • v.36 no.2
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    • pp.140-146
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    • 2011
  • This study was performed to develop a rapid test kit for pathogenic Salmonella in various samples. The rapid detection kit has been fabricated based on nitrocellulose lateral-flow strip. Colloidal gold and biotin conjugated Salmonella antibodies were used as a tag and a receptor, respectively. Manually spotted Salmonella antibody and Neutravidin on nitrocellulose membrane were used as test and control lines, respectively. Feasibility of the rapid kit to detect Salmonella typhimurium in samples were evaluated. The intensity of the color of the test line started to increase with the samples in which higher concentration of the cells were contained. The sensitivity of the sensor was $10^6$ cfu/mL Salmonella spiked in PBS. Also, the rapid test kit could detect $10^6$ cfu/mL of Salmonella in chicken meat extract.

A Design of An Active PID control using Immune Algorithm for Vibration Control of Building Structure (구조물 진동제어를 위한 Immune Algorithm을 이용한 Active PID 제어기 설계)

  • Lee, Young-Jin;Cho, Hyun-Cheol;Lee, Kwon-Soon
    • Proceedings of the KIEE Conference
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    • 2005.05a
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    • pp.72-74
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    • 2005
  • In this paper, we propose an adaptive PID controller using a cell-mediated immune response to improve a PID control performance. The proposed controller is based on the specific immune response of the biological immune system that is cell-mediated immunity. The immune system of organisms in the real body regulates the antibody and the T-cells to protect an attack from the foreign materials like virus, germ cells, and other antigens. It has similar characteristics that are the adaptation and robustness to overcome disturbances and to control the plant of engineering application. We first build a model of the T-cell regulated immune response mechanism and then designed an I-PID controller focusing on the T-cell regulated immune response of the biological immune system. We apply the proposed methodology to building structures to mitigate vibrations due to strong winds for evaluation of control performances. Through computer simulations, system responses are illustrated and additionally compared to traditional control approaches.

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