• 제목/요약/키워드: Antibody Engineering

검색결과 349건 처리시간 0.029초

Surface Plasmon Resonance Imaging Analysis of Hexahistidine-tagged Protein on the Gold Thin Film Coated with a Calix Crown Derivative

  • Chung, Bong-Hyun;Baek, Seung-Hak;Shin, Yong-Beom;Kim, Min-Gon;Ro, Hyeon-Su;Kim, Eun-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권2호
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    • pp.143-146
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    • 2004
  • A surface plasmon resonance (SPR) imaging system was constructed and used to detect the hexahistidine-ubiquitin-tagged human parathyroid hormone fragment (His$\sub$6/-Ub-hPTHF(1-34)) expressed in Escherichia coli. The hexahistidine-specific antibody was immobilized on a thin gold film coated with ProLinker$\^$TM/ B, a novel calixcrown derivative with a bifunctional coupling property that permits efficient immobilizaton of capture proteins on solid matrices. The soluble and insoluble fractions of an E. coli cell lysate were spotted onto the antibody-coated gold chip, which was then washed with buffer (pH 7.4) solution and dried. SPR imaging measurements were carried out to detect the expressed His$\sub$6/-Ub-hPTHF(1-34). There was no discernible protein image in the uninduced cell lysate, indicating that non-specific binding of contaminant proteins did not occur on the gold chip surface. It is expected that the approach used here to detect affinity-tagged recombinant proteins using an SPR imaging technique could be used as a powerful tool for the analyses of a number of proteins in a high-throughput mode.

Hep88 mAb-Mediated Paraptosis-Like Apoptosis in HepG2 Cells via Downstream Upregulation and Activation of Caspase-3, Caspase-8 and Caspase-9

  • Mitupatum, Thantip;Aree, Kalaya;Kittisenachai, Suthathip;Roytrakul, Sittiruk;Puthong, Songchan;Kangsadalampai, Sasichai;Rojpibulstit, Panadda
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권5호
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    • pp.1771-1779
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    • 2015
  • Hepatocellular carcinoma (HCC) is a leading cause of cancer death worldwide. Presently, targeted therapy via monoclonal antibodies to specific tumor-associated antigens is being continuously developed. Hep88 mAb has proven to exert tumoricidal effects on the HepG2 cell via a paraptosis-like morphology. To verify the pathway, we then demonstrated downstream up-regulation of caspase-3, caspase-8 and caspase-9, assessingmRNA expression by real-time PCR and associated enzyme activity by colorimetric assay. Active caspase-3 determination was also accomplished by flow cytometry. Active caspase-3 expression was increased by Hep88 mAb treatment in a dose-and time-dependent manner. All of the results indicated that Hep88 mAb induced programmed cell death in the HepG2 cell line from paraptosis-like to apoptosis by downstream induction of caspases. These conclusions imply that Hep88mAb might be a promising tool for the effective treatment of HCC in the future.

Effects of Gal-13 on the Content of Immunoglobulin, Proliferation of Lymphocyte and Antibody Titers after Vaccination with Infectious Bursal Disease Virus Vaccine in Chickens

  • Yang, Yurong;Jiang, Yibao;She, Ruiping;Peng, Kaisong;Zhou, Xuemei;Yin, Qingqiang;Wang, Decheng;Liu, Tianlong;Bao, Huihui
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권3호
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    • pp.405-411
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    • 2007
  • Gal-13 is an antimicrobial peptide isolated from chicken intestine. Ninety chickens were randomly divided into two groups (45 chickens for each group) to determine the effect of oral administration of Gal-13 on the acquired immune response. The chickens in the first group were fed a diet without Gal-13 as the control, and the chickens in the second group were fed the same diet, except that Gal-13 ($1{\mu}g/ml$) was suspended in drinking water just after hatching. Samples of blood, thymus, bursa of fabricius and spleen were taken at day 1, 4, 7, 10 and 17. The chickens in both groups received infectious bursal disease virus vaccine at day 20, and then sera samples were collected for analysis at 14, 21, 28 and 35 days after vaccination. The results showed: (1) Gal-13 could enhance the content of immunoglobulin (Ig)G at the age of 4 to10 days (p<0.05) and IgM at the age of 4 and 10 days (p<0.05) in the serum; (2) In vitro experiments showed that Gal-13 (0.625-1.250${\mu}g/ml$) enhanced the proliferation of peripheral blood lymphocytes of the chickens stimulated by lipopolysaccharide (LPS) and concanavlin A (ConA). Compared to the control, Gal-13 (1 ${\mu}g/ml$) enhanced the proliferation of bursa lymphocytes at 17 days of age (p<0.01) and thymus lymphocytes at 7 days of age (p<0.01), but restrained lymphocyte proliferation in chicken spleen and differed significantly at day 10 (p<0.01); (3) Gal-13 enhanced infectious bursal disease virus antibody in sera of chickens 21 days after infectious bursal disease virus vaccine administration (p<0.05). These results suggested that Gal-13 could modulate adaptive immune responses of chickens.

살균제 Iprovalicarb 잔류물의 신속한 검출을 위한 바이오센서 (A Biosensor for the Rapid Detection of the Fungicide Iprovalicarb Residuess)

  • 조한근;김운호;경기성;이은영
    • Journal of Biosystems Engineering
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    • 제32권6호
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    • pp.440-447
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    • 2007
  • In this study, a biosensor was developed using an enzyme-linked immunosorbent assay (ELISA) to rapidly measure the fungicide iprovalicarb residues in agricultural products. The biosensor was designed to include micro-pumps and solenoid valves for fluid transport, a spectrophotometer cuvet as a reaction chamber, a photodiode with a light-emitting diode for optical density measurement, and a control microcomputer to implement assay. The rate of change in optical density of the cuvet was read as final signal output. Micro-pumps were evaluated to investigate their delivery capability, the highest values of the error and the coefficient of variation were 4.3% and 4.6% respectively. As the incubation period was reduced from 15 minutes to 11 minutes to shorten the total processing time, the sensor sensitivity was decreased as the antibody dilution ratio was reduced to a half. The maximum usable period of the coated cuvet was found to be two days with 1% error limit. To predict the concentration of the iprovalicarb residue in agricultural products, a linear calibration model was obtained with r-square values of 0.992 for potato and 0.985 for onion. In validation test for the samples of potatoes and onions against the high performance liquid chromatography, very high correlation values were obtained as 0.996 and 0.993 respectively. Using the cuvet immobilized with antigen, it took 21-minutes for the biosensor to complete the measuring process of the iprovalicarb residues.

노로바이러스 검출을 위한 측면유동면역분석법 기반의 바이오리셉터 선별기법 개발 (Norovirus Targeted Bioreceptor Screening Method based on Lateral Flow Immunoassay (LFIA))

  • 장희수;조현지;전태준;김선민
    • 한국가시화정보학회지
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    • 제20권3호
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    • pp.136-145
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    • 2022
  • Later flow immunoassay (LFIA) is a protein analytical method based on immunoreaction. On the LFIA based protein analytical method, bioreceptor molecule plays a key role, and so a system that evaluates and manages the binding affinity of bioreceptor is needed to secure detection reliability. In this study, Lateral Flow Immunoassay based rapid Bioreceptor Screening Method (rBSM) is presented that provide a simple and quick evaluating method for the binding affinity to the target protein of the antibody as model bioreceptor. To verify this evaluation method, Virus-like particles (VLP) and anti-VLP antibodies are selected as a model norovirus, which is target protein, and the candidate bioreceptors respectively. Among the 5 different candidate antibodies, appropriate antibody could be sorted out within 30 minutes through rBSM. In addition, selected antibodies were applied to two representative LFIA based techniques, sandwich assay and competitive assay. Among these methods, sandwich assay showed more effective VLP detection method. Through applying selected antibodies and techniques to the commercialized mass production lines, an VLP detecting LFIA kit was developed with a detection limit of 1012 copies/g of VLPs in real samples. Since this proposed method in this study could be easily transformable into other combinations with bioreceptors, it is expected that this technique would be applied to LFIA kit development system and bioreceptor quality management.

Development of Chemiluminescent Immunosensor Array for GMO

  • Jung, Woo-Sung;Hwang, Ok-Hwa;Jang, Hye-Ji;Paek, Eui-Hwan;Park, Won-Mok;Paek, Se-Hwan
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
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    • pp.683-686
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    • 2003
  • While genetically modified organisms (GMOs) are producing in many countries, issues related to safeties of GMOs as foods for human have risen. Because of such potential problems, the development of an indication system regarding GMO content contained in foods has been required. Particularly, an immune-chip, as widely demanded diagnostic tool for functional, structural analyses of proteins, has been investigated to simultaneously measure different analytes. We have developed methods for immobilizing antibody on glass surfaces as substrate and for generating chemiluminometric signals.

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항체공학 (Antibody Engineering)

  • 홍효정
    • 미생물과산업
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    • 제17권1호
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    • pp.51-55
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    • 1991
  • 쥐의 단일클론항체는 기초연구에 뿐만 아니라 질병의 진단시약으로서도 광범위하게 유용되어 왔다. 반면, 쥐의 단일클론항체를 인체에 임상적으로 사용하는 것은 인체에 면역반응을 유발시킨다는 근본적인 문제점 때문에 어려운 일이라 인식되어 왔다. 그러나, 최근 2-3년간 유전자 기술과 단백질공학적인 방법에 의하여 쥐의 단일클론항체를 인간화시키거나 항체절편의 생산을 시도하게 되어, 단일클론항체의 임상적인 이용에 대한 가능성이 긍정적으로 진단되고 있다. 본 난에서는 항체의 구조와 기능을 간략히 살펴보고, 최근 항체의 단백질공학에 대한 경향과 방법 및 engineered 항체의 특성들에 관하여 논의하고자 한다.

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