• 제목/요약/키워드: avidin-biotin

검색결과 102건 처리시간 0.029초

Synthesis and Characterization of a Receptor-Targeting Contrast Agent

  • Yang, Taegyun;Park, Ji-Hyung;Lee, Seung-Cheol;Kim, Cheol-Su;Cho, Jee-Hyun;Lee, Chul-Hyun;Cheong, Chae-Joon
    • 한국자기공명학회논문지
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    • 제7권1호
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    • pp.46-54
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    • 2003
  • We synthesized a contrast agent for MRI that is capable of binding to the ABP-1 receptor and enhancing the contrast of the targeted cells. We used a lysine dendrimer (G=3)DTPA[Gd] as the contrast agent and synthesized a biotinylated polyclonal antibody for ABP-1 as the first antibody. Lysine dendrimers were prepared using the solid phase peptide synthesis method.$^3$ Amino-terminated lysine dendrimers were then coupled to DTPA using the anhydride method. Gd was complexed with the DTPA-lysine dendrimer in an acidic solution of 3 eq GdCl$_3$ to one of DTPA. The lysine dendrimer-DTPA[Gd] and avidin were conjugated in MES solution, pH 6.0, using EDC as the coupling reagent. The biotin-avidin system was used to link the polyclonal antibody and contrast agent. K562 cells were used for imaging.

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제초제 검출을 위한 전기화학적 일회용 면역센서 (Disposable Electrochemical Immunosensors for the Detection of Herbicide)

  • 장승철
    • 센서학회지
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    • 제20권1호
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    • pp.35-39
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    • 2011
  • A disposable electrochemical immunosensor system has been developed for the detection of herbicide in aqueous samples. Disposable screen printed carbon electrodes(SPCE) were used as basic electrodes and an enzyme, horseradish peroxidase (HRP), and anti-herbicide antibodies was immobilised on to the working electrode of SPCE by using avidin-biotin coupling reactions. An herbicide-glucose oxidase conjugates have been used for the competitive immunoreaction with sample herbicides. The enzymatic reaction between the conjugated glucose oxidase and glucose added generates hydrogen peroxide, which was reduced by the peroxidase immobilised. The latter process caused an electrical current change, due to direct re-reduction of peroxidase by a direct electron transfer mechanism, which was measured to determine the herbicides in the sample. The optimal operational condition was found to be: $20\;{\mu}gl-1$ deglycosylated avidin loading to the working electrode and working potential +50 mV vs. Ag/AgCl. The total assay time was 15 min after sample addition. The detection limits for herbicides, atrazine and simazine, were found to be 3 ppb and 10 ppb, respectively.

Efficient Biotinylation of Nitrocellulose Membrane for Immuno-Filtration Capture Assay

  • Choi, Ki-Bong;Ha, Youn-Chul;Youn, Hee-Ju;Choi, Jung-Do
    • BMB Reports
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    • 제30권5호
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    • pp.308-314
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    • 1997
  • We investigated biotinylation of nitrocellulose membrane for immuno-filtration capture assay. In order to enhance the efficiency of biotinylation, nitrocellulose membranes were pretreated with several chemicals for the purpose of suitable protein absorption through surface modification. As a signal generating enzyme, urease was used and the concentration of avidin was optimized for the efficient binding kinetics between urease-biotin in liquid phase and biotinylated membrane in solid phase. For effective biotinylation, bovine serum albumin-biotin complexes could be immobilized at a concentration of $370\;{\mu}g$/stick ($4.4\;cm^2$). Among tested chemicals, polylysine (0.25%) showed a significant effect in biotinylation. Polylysine is thought to enhance surface area by extending unbound residues into solution. Time of treatment over 30 min and higher molecular weight of polylysines (58,100 dalton) showed positive effect on the enhancement of biotinylation. The result from this study may be useful for developing a new biosensor and other biofunctional membranes for examining molecular recognition.

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서로 다른 두 작용기를 이용한 결합법에 의한 접합체: 도파민 면역분석법 (Bioconjugation by dual heterobifunctional coupling method: Use of the conjugates for the detection of dopamine)

  • 류지은;이인숙
    • 분석과학
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    • 제23권6호
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    • pp.537-543
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    • 2010
  • 도파민은 카테콜아민류의 중요한 신경전달물질로서 부족하면 파킨스병과 정신분열증 등을 야기할 수 있다. 그러므로 조작이 비교적 간단하면서 감도가 우수한 분석법의 개발이 필요하다. 이에, 도파민에 대한 경쟁적인 효소면역분석법이 연구되었다. 경쟁적인 면역분석법의 분석감도는 일반적으로 두가지 요소에 의해 조절된다. 하나는 경쟁자의 특성과 농도이며, 다른 하나는 결합체, 즉 항체의 그것이다. 따라서, 경쟁자인 BSA-DA과 결합체인 항체-avidin 접합체의 최적화가 수행되었다. 두 접합체는 SATA와 SMCC를 이용한 dual heterobifunctional coupling법에 의해 합성되었으며, 최적화 과정을 통해 BSA-DA 접합체의 농도는 $6.66\;{\mu}g/mL$, 항체-avidin 접합체의 농도 $4.17{\times}10^{-10}\;M$로 결정되었다. 도파민에 대한 doseresponse curve와 calibration curve의 결과로써 도파민에 대한 검출 한계는 $2.3{\times}10^{-2}\;{\mu}g/mL$ 이고 검출 영역은 $1.0{\times}10^{-3}\;M\sim1.0{\times}10^{-7}\;M$ 이다. 직선성을 갖는 검출영역에서의 검정선을 얻은 결과 [Absorbance = -0.1098 log[DA]+0.0353 ($R^2$ = 0.9956)] 우수한 직선관계를 얻었다.

기능성 고분자의 세포특이성 재료로의 이용에 관한 연구 (Application of Hepatocyte Specific Polymers with Functional group)

  • 이정복;김재웅
    • 분석과학
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    • 제9권1호
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    • pp.84-90
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    • 1996
  • 본 연구에서는 스티렌 골격에 올리고당 및 비오틴 잔기를 결사슬에 갖는 새로운 세포 특이성 고분자 재료를 설계하여 합성하였다. 이들 고분자 재료를 간세포의 다점 인식 재료로서의 사용 가능성을 검토한 실험 결과를 보고하고자 한다. 간세포의 초기 접착 거동을 조사하기 위하여 합성된 고분자 재료 p(VLA-co- VBA) 90 : 10, p(VLA-co-VBA) 80 : 20 및 대조 물질로 PVLA의 농도가 각각 0.01%(w/v) 되도록 조절하여 1mL 씩 폴리스티렌 페트리 접시에 넣고, 혈청 존재 및 비존재하에서 Seglen법으로 단리한 간세포를 각각 첨가한 결과 60분 경과 후에는 혈청 비존재하에서와 같은 70% 정도의 높은 접착률을 보여 주었다. 폴리머 p(VLA-co-VBA) 70:30에 함유된 비오틴 잔기와 아비딘과의 분자 회합에 의한 응집형상은 UV 투과율의 변화로 확인 하였다.

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알츠하이머 조기 진단을 위한 변형된 대식세포의 기초적 연구 (Primary Cellular Study of Phagocytosis for Alzheimer Disease Diagnosis)

  • 조정민;채철주;강재민;김관수;송기봉
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2010년도 하계학술대회 논문집
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    • pp.280-280
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    • 2010
  • Alzheimer disease is a progressive neurodegenerative disease of the aged, characterized by memory loss and dementia. For diagnosis of Alzheimer disease we have simply modified macrophage with amyloid beta bonded with different molecules. Modified Macrophage was observed with microscope for co-localization of amyloid beta molecule. For this experiment we used fluoroscene labeling substances. The macrophage was modified also with cell staining method. For cell staining method was used avidin-biotin reaction principles. All experiments were carried out on poly-L-lysine coated and sterilized glass substrates. In the presentation we will show the further investigations and applications with modified macrophage.

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치주낭 조직내 tenascin의 분포에 관한 면역조직화학적 연구 (AN IMMUNOHISTOCHEMICAL LOCALIZATION OF TENASCIN IN PERIODONTAL POCKET TISSUES)

  • 한경윤;이강진
    • Journal of Periodontal and Implant Science
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    • 제24권3호
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    • pp.607-617
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    • 1994
  • To determine the effect of tenascin on forming periodontal pocket and pseudopocket, the ginival tissues were surgically obtained from the patients with adult periodontitis(10) and non-inflammatory phenytoin-associated gingival hyperplasia(5). The excised tissue specimens were fixed in neutral formalin for $6{\sim}24$ hours, embedded with paraffin, sectioned at 4-6m in thickness, mounted on glass slides coated with 3-aminopropyltriethoxysilane(Sigma Chemical Co., St. Louis, MO, U.SA.) and immunohistochemically processed by Avidin-Biotin peroxidase complex method for the localization of tenascin, using monoclonal mouse anti-human tenascin antiboday(Chemicon-International Inc., Temecula, CA, U.S.A., 1: 5,000) as the primary antibody. Regardless of periodontal pocket and pseudopocket, tenascin was localized along the connective tissue subjacent to basement membrane of gingival epithelium, and strong positive reactivity was obviously noted in the papillary projections of gingival connective tissue. The results suggest that tenascin may affect the development of papillary projections and the proliferation of epithelial cells.

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유아기 흑색 신경외배엽성 종양의 치험례 (MELANOTIC NEUROECTODERMAL TUMOR OF INFANCY ; A CASE REPORT)

  • 김일규;하수용;이성준;주영채
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제13권4호
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    • pp.436-443
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    • 1991
  • 저자등은 인하대학교 부속병원에 내원한 생후 5개월된 여아의 상악 전치부에서 발생한 MNTI 1례를 en-bloc excision으로 치험하고, 면역조직화학을 이용한 염색으로 MNTI가 신경능에서 기원하였음을 확인할 수 있었다.

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Preparation and Atomic Force Microscopy (AFM) Characterization of DNA Scaffolds as a Template for Protein Immobilization

  • Kim, Hyeran;Lee, Hyun Uk;Lee, Jouhahn
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2014년도 제46회 동계 정기학술대회 초록집
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    • pp.411.2-411.2
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    • 2014
  • The design of DNA nanostructures is of fundamental importance, the intrinsic value of DNA as a building-block material lies in its ability to organize other bio-molecules with nanometer-scale spacing. Here, we report the fabrication of DNA scaffolds with nano-pores (<10 nm size) that formed easily without the use of additives (i.e., avidin, biotin, polyamine, or inorganic materials) into large-scale structures by assembling DNA molecules at near room temperature ($30^{\circ}C$) and low pH (~5.5). Protein immobilization results also confirmed that a fibronectin (FN) proteins/large scale DNA scaffolds/aminopropylytriethoxysilane (APS)/SiO2/Si substrate with high sensitivity formed in a well-defined manner. The DNA scaffolds can be applied for use with DNA-based biochips, biophysics, and cell biology.

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