• Title/Summary/Keyword: antibody engineering

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Investigation of the Binding Force between Protein A and Immunoglobulin G Using Dielectrophoretic(DEP) Tweezers Inside a Microfluidic Chip (미세유체 칩 내에서 유전영동 집게(Dielectrophoretic Tweezers) 를 이용한 단백질A와 면역 글로불린 G의 결합에 관한 연구)

  • Kwak, Tae Joon;Lee, Jae Woo;Yoon, Dae Sung;Lee, Sang Woo
    • Journal of Biomedical Engineering Research
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    • v.34 no.3
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    • pp.123-128
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    • 2013
  • The 'Dielectrophoretic Tweezers(DEP Tweezers)' can be used as a facile, economical toolkit for quantitative measurement of chemical and biological binding forces related to many biological interactions within a microfluidic device. Our experimental setup can probe the interaction between a single receptor molecule and its specific ligand. Immunoglobulin G(IgG) functionalized on polystyrene microspheres has been used to detect individual surface linked Staphylococcus protein A(SpA) molecules and to characterize the strength of the noncovalent IgG-SpA bond. It was measured and compared with the existing measurements. Measured single binding force of between Goat, Rabbit IgG and SpA were $17{\pm}7pN$, $74{\pm}16pN$. This work can be used to investigate several different ligand-receptor interactions and antigen-antibody interactions.

Active Distribution System Planning Considering Battery Swapping Station for Low-carbon Objective using Immune Binary Firefly Algorithm

  • Shi, Ji-Ying;Li, Ya-Jing;Xue, Fei;Ling, Le-Tao;Liu, Wen-An;Yuan, Da-Ling;Yang, Ting
    • Journal of Electrical Engineering and Technology
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    • v.13 no.2
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    • pp.580-590
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    • 2018
  • Active distribution system (ADS) considering distributed generation (DG) and electric vehicle (EV) is an effective way to cut carbon emission and improve system benefits. ADS is an evolving, complex and uncertain system, thus comprehensive model and effective optimization algorithms are needed. Battery swapping station (BSS) for EV service is an essential type of flexible load (FL). This paper establishes ADS planning model considering BSS firstly for the minimization of total cost including feeder investment, operation and maintenance, net loss and carbon tax. Meanwhile, immune binary firefly algorithm (IBFA) is proposed to optimize ADS planning. Firefly algorithm (FA) is a novel intelligent algorithm with simple structure and good convergence. By involving biological immune system into FA, IBFA adjusts antibody population scale to increase diversity and global search capability. To validate proposed algorithm, IBFA is compared with particle swarm optimization (PSO) algorithm on IEEE 39-bus system. The results prove that IBFA performs better than PSO in global search and convergence in ADS planning.

Effects of Dietary Chromium Picolinate Supplementation on Growth Performance and Immune Responses of Broilers

  • Lee, Der-Nan;Wu, Fu-Yu;Cheng, Yeong-Hsiang;Lin, Rong-Shinn;Wu, Po-Ching
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.2
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    • pp.227-233
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    • 2003
  • Two experiments were conducted to evaluate the effects of chromium (Cr) on the growth performance, bone trait, serum traits, and immune responses in broilers. The broilers were fed corn-soybean meal basal diet supplemented with Cr at level of 0(control), 200, 400, or 800 ppb in the form of chromium picolinate (CrPic). The broilers were fed treated diets for 6 weeks in Exp. 1, but the Cr supplement was removed for the last 3 weeks in Exp. 2. Exp. 1 showed that dietary supplement of Cr did not affect growth performance of the broiler, though improved feed efficiency (p<0.05) was observed during 0 to 3 weeks. Moreover, serum total (p<0.05) and HDL cholesterols (p<0.06) were significantly higher in pooled Cr added group at 6 weeks of age, however, the difference was not significant in Exp. 2. The pooled Cr added group in Exp.1 had significantly lower (p<0.05) alkaline phosphatase activity and higher (p<0.09) calcium at 3 weeks. Significantly lower phosphorus was also observed in Exp. 2. With continued supplement of Cr as in Exp. 1, the alkaline phosphatase activity maintained higher at 6 weeks, as opposed to significantly lower in Exp. 2, which had no further Cr supplement. Higher bone breaking strength was observed in 400 ppb Cr supplemented in Exp. 1, though not significantly different. Serum glucose and triglyceride were not affected by Cr supplement. Antibody against Infectious Bronchitis (IB) was significantly (p<0.05) higher with 400 ppb Cr supplemented, and anti-Newcastle disease (ND) antibody also tended to be higher (p<0.06) in pooled Cr added group at 6 weeks of age in Exp. 1. Peripheral blood blastogenesis activity was not different among the treatments. The results suggest that diet supplemented with 400 ppb CrPic may be beneficial to the broiler.

Expression of the Apx Toxins of Actinobacillus pleuropneumoniae in Saccharomyces cerevisiae and Its Induction of Immune Response in Mice

  • Park Seung-Moon;Choi Eun-Jin;Kwon Tae-Ho;Jang Yong-Suk;Yoo Han-Sang;Choi Woo Bong;Park Bong-Kyun;Kim Dae-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.362-366
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    • 2005
  • Actinobacillus pleuropneumoniae is an important pig pathogen, which is responsible for swine pleuropneumonia, a highly contagious respiratory infection. To develop subunit vaccines for A. pleuropneumoniae infection, the Apx toxin genes, apxI and apxII, which are thought to be important for protective immunity, were expressed in Saccharomyces cerevisiae, and the induction of immune responses in mice was examined. The apxI and apxII genes were placed under the control of a yeast hybrid ADH2-GPD promoter (AG), consisting of alcohol dehydrogenase II (ADH2) and the GPD promoter. Western blot analysis confirmed that both toxins were successfully expressed in the yeast. The ApxIA and ApxIIA-specific IgG antibody response assays showed dose dependent increases in the antigen-specific IgG antibody titers. The challenge test revealed that ninety percent of the mice immunized with ApxIIA or a mixture of ApxIA and ApxIIA, and sixty percent of mice immunized with ApxIA survived, while none of those in the control groups survived longer than 36 h. These results suggest that vaccination of the yeast ex­pressing the ApxI and ApxII antigens is effective for the induction of protective immune responses against A. pleuropneumoniae infections in mice.

Isolation of Mouse Ig Heavy and Light Chain Genomic DNA Clones, and Construction of Gene Knockout Vector for the Generation of Humanized Xenomouse (인간 단클론 항체 생산용 Humanized Xenomouse 제작의 기초 소재인 생쥐 Ig 중사슬 및 경사슬 Genomic DNA 클론의 확보 및 유전자 적중 벡터의 제작)

  • Lee, Hee-kyung;Cha, Sang-hoon
    • IMMUNE NETWORK
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    • v.2 no.4
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    • pp.233-241
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    • 2002
  • Background: Monoclonal antibodies (mAb) of rodent origin are produced with ease by hybridoma fusion technique, and have been successfully used as therapeutic reagents for humans after humanization by genetic engineering. However, utilization of these antibodies for therapeutic purpose has been limited by the fact that they act as immunogens in human body causing undesired side effects. So far, there have been several attempts to produce human mAbs for effective in vivo diagnostic or therapeutic reagents including the use of humanized xenomouse that is generated by mating knockout mice which lost Ig heavy and light chain genes by homologous recombination and transgenic mice having both human Ig heavy and light gene loci in their genome. Methods: Genomic DNA fragments of mouse Ig heavy and light chain were obtained from a mouse brain ${\lambda}$ genomic library by PCR screening and cloned into a targeting vector with ultimate goal of generating Ig knockout mouse. Results: Through PCR screening of the genomic library, three heavy chain and three light chain Ig gene fragments were identified, and restriction map of one of the heavy chain gene fragments was determined. Then heavy chain Ig gene fragments were subcloned into a targeting vector. The resulting construct was introduced into embryonic stem cells. Antibiotic selection of transfected cells is under the progress. Conclusion: Generation of xenomouse is particularly important in medical biotechnology. However, this goal is not easily achieved due to the technical difficulties as well as huge financial expenses. Although we are in the early stage of a long-term project, our results, at least, partially contribute the successful generation of humanized xenomouse in Korea.

Improvement of Functional Properties of Egg White Protein through Glycation and Phosphorylation by Dry-heating

  • Enomoto, Hirofumi;Nagae, Shiho;Hayashi, Yoko;Li, Can-Peng;Ibrahim, Hisham R.;Sugimoto, Yasushi;Aoki, Takayoshi
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.4
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    • pp.591-597
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    • 2009
  • Egg white protein (EWP) was glycated with maltopentaose (MP) through the Maillard reaction and subsequently phosphorylated by $85^{\circ}C$ dry-heating at pH 4.0 for 1 d in the presence of pyrophosphate. The functional properties of glycated, phosphorylated EWP were compared with those of native EWP and with EWP which was phosphorylated by dry-heating in the presence of pyrophosphate under the same conditions. The phosphorus content of EWP was increased to ~0.60% by phosphorylation, and to ~0.74% by glycation with MP and subsequent phosphorylation. The electrophoretic mobility of EWP increased through phosphorylation. The stability of EWP against heat-induced insolubility at pH 7.0 was considerably improved by phosphorylation alone and further by phosphorylation after glycation. The anti-ovalbumin antibody response was reduced significantly by glycation and phosphorylation, and further reduced by phosphorylation after glycation. The anti-ovomucoid antibody response was reduced significantly by glycation, phosphorylation and phosphorylation after glycation. The calcium phosphate-solubilizing ability of EWP was enhanced by both phosphorylation methods.

Image Analysis of a Lateral Flow Strip Sensor for the Detection of Escherichia coli O157:H7

  • Kim, Giyoung;Moon, Ji-Hea;Park, Saet Byeol;Jang, Youn-Jung;Lim, Jongguk;Mo, Changyeun
    • Journal of Biosystems Engineering
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    • v.38 no.4
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    • pp.335-340
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    • 2013
  • Purpose: This study was performed to develop a lateral flow strip sensor for the detection of pathogenic Escherichia coli O157:H7 in various samples. Also, feasibility of using an image analysis method to improve the interpretation of the strip sensor was evaluated. Methods: The lateral flow strip sensor has been fabricated based on nitrocellulose lateral-flow membrane. Colloidal gold and E. coli O157:H7 antibodies were used as a tag and a receptor, respectively. Manually spotted E. coli O157:H7 antibody and anti-mouse antibody on nitrocellulose membrane were used as test and control dots, respectively. Feasibility of the lateral flow strip sensor to detect E. coli O157:H7 were evaluated with serially diluted E. coli O157:H7 cells in PBS or food samples. Test results of the lateral flow strip sensor were measured with an image analysis method. Results: The intensity of the test dot started to increase with higher concentration of the cells were introduced. The sensitivities of the sensor were both $10^4$ CFU/mL Escherichia coli O157:H7 spiked in PBS and in chicken meat extract, respectively. Conclusions: The lateral flow strip sensor and image analysis method could detect E. coli O157:H7 in 20 min, which is significantly quicker than conventional plate counting method.

Generation of myelination with neural cell cultures in rats and suppression of myelination by infection of sindbis virus (쥐의 신경세포 배양에 의한 수초 발생과 sindbis 바이러스 감염에 의한 수초 억제)

  • Sa, Young-Hee;Kim, Hyun Joo;Lee, Bae Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2019.05a
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    • pp.528-532
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    • 2019
  • The dorsal root ganglion (DRG) was isolated from mouse embryos and Schwann cells and neuronal cells were cultured in vitro. The neurons and Schwann cells were cultured separately and the two kinds of cells were cultured together for three weeks. Generation of myelination was confirmed by transmission electron microscope and confocal microscope using a myelinaion protein, myelin protein zero (MPZ) antibody. The sindbis virus was infected for three days in the myelinated culture cells and then demyelination was carried out. The process of demyelination was also confirmed by transmission electron microscopy and confocal microscopy using myelin protein zero (MPZ) antibody. The study was supported by a Basic Research Program through the National Research Foundation (NRF) funded by the Ministry of Science and Technology, ICT and Future Plans (NRF-2016R1A2B4016552 and 2017R1A2B3005753).

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Analysis of the influence degree of each factor on the linkage affecting the lever actuating force in an implant transport device for the treatment of eye diseases (안과질환 처치를 위한 임플란트 수송장치에서 레버 작동력에 영향을 주는 연동장치에 대한 인자별 영향도 분석 )

  • Jeong-Won Lee;Joong-Seob Guk
    • Design & Manufacturing
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    • v.18 no.3
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    • pp.1-8
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    • 2024
  • Macular degeneration is a disease that damages the macula, the center of the retina, and is one of the three major eye diseases along with glaucoma and diabetic retinopathy. The optic nerve and most of the photoreceptor cells are located here, and since this is where images of objects are formed, it is the most important area for vision. The main symptom of macular degeneration is the inability to clearly distinguish the shape of objects or the inability to distinguish colors and light and dark. It is also a serious eye disease that causes black spots in the center of the field of vision. However, it is difficult to distinguish it from the form of vision loss due to presbyopia, so early diagnosis is often missed. The most common treatment for macular degeneration is antibody injection therapy. This treatment requires regular injections once every 1-2 months. When receiving antibody injection therapy, the fear of having to inject directly into the eye and the cost of long-term repeated procedures are a great burden to patients. To overcome these problems, special sustained-release formulations using drug delivery systems are being developed. Since the release speed and release time of the drug can be controlled, the number of times the drug is administered can be drastically reduced. However, the implant (Ø 0.46×6.0mm), which is a sustained-release agent, is manufactured by mixing biodegradable resin (PLGA) and therapeutic agent in a ratio of 4:6, so it is very brittle and there is a high risk of implant damage during handling. In order to safely insert the implant into the eye, a transport device that can be driven with controlled force is required. Therefore, in this study, the lever operating force was measured and analyzed to determine the influence of factors according to the cross-sectional thickness and shape of the linkage produced through injection molding as well as the post-process.

Single-Step Purification of Proteins of Interest from Proteolytically Cleaved Recombinant Maltose-binding Protein (MBP) Fusion Proteins by Selective Immunoprecipitation of MBP

  • Park, Jung-Hyun;Na, Shin-Young;Lee, Dong-Gun;Han, Byoung-Don;Kim, Kil-Lyong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.82-86
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    • 1998
  • The maltose binding protein (MBP) fusion protein system is a versatile tool to express and isolate recombinant proteins in E. coli. In this system, MBP fusion proteins are efficiently isolated from whole cell lysate using amylose conjugated agarose beads and then eluted by competition with free maltose. Since MBP is a rather large molecule (∼42 kDa), for further experiments, the MBP part is usually proteolytically cleaved from the fusion protein and subsequently removed by ion-exchange chromatography or rebinding to amylose columns after washing out excess and MBP-bound maltose. In the present study, we have developed an improved method for the removal of cleaved MBP, which is advantageous over conventional methods. In this method, factor Xa cleaved MBP fusion proteins were incubated with Sepharose beads conjugated with MBP specific monoclonal antibodies and then precipitated buy centrifugation, resulting in highly purified proteins in the supernatant.

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