• 제목/요약/키워드: Polyclonal antibody

검색결과 283건 처리시간 0.032초

Production rind Characterization of the Polyclonal Anti-peptide Antibody for $\beta$-adrenergic Receptor

  • Kim, Hee-Jin;Shin, Chan-Young;Sang Bong lee;Ko, Kwang-Ho
    • Biomolecules & Therapeutics
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    • 제2권4호
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    • pp.303-309
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    • 1994
  • The analysis of membrane receptors for hormones and neurotransmitters has progressed considerably by pharmacological and biochemical means and more recently through the use of specific antibodies. Two kinds of antibodies could be produced, one is from synthetic peptides and the other from proteins such as purified receptor. Anti-peptide antibodies gave some advantages; epitope is evident and also receptor purification in quantity is not prerequisite. It can be also applied to the study of receptor structure-activity relationship. The purpose of the present study was 1) to produce and characterize a polyclonal antibody against a synthetic $\beta$2-adrenergic receptor peptide(Phe-Gly-Asn-Phe-Trp-Cys-Phe-Trp-Thr-Ser-Ile-Asp-Val-Leu) and 2) to determine the effects of this antibody on the $\beta$-adrenergic receptor ligand interaction. The peptide sequence contains an amino acid residue such as Asp-113 which was identified as one of important component for receptor-ligand interaction in site-directed mutagenesis studies. Production of antibody was performed by immunization of rabbits through popliteal lymph node with the peptide coupled with Keyhole Limpet Hemocyanin (KLH). The titer of antibody against this peptide was 1 : 1000. The anti-peptide antibody was able to detect a 67 kDa protein band in western blot corresponding to the molecular weight of the $\beta$-adrenergic receptor in partially purified receptor fraction derived from guinea pig lung. The antisera inhibited the specific binding of [$^3$H]dihydroalprenolol to $\beta$-adrenergic receptor in a concentration-dependent manner. The results from this study suggest that the peptide sequence selected in the present study is important for the receptor ligand interaction.

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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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Purification, Partial Characterization, and Immunoassay of Vitellogenin from Marbled Sole (Limanda yokohamae)

  • Kim Dae-Jung;Jung Jee-Hyun;An Cheul-Min;Jee Young-Ju;Min Kwang-Sik;Kim Yoon;Han Chang-Hee
    • Fisheries and Aquatic Sciences
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    • 제8권4호
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    • pp.213-219
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    • 2005
  • Vitellogenin (VTG) was purified from the blood plasma of estradiol-17$\beta$ ($E_2$)-treated male marbled sole (Limanda yokohamae) using gel filtration and anion exchange chromatography. The purity of the marbled sole VTG (msVTG) was confirmed by polyacrylamide gel electrophoresis (SDS-PAGE) and N-terminal amino acid sequencing. The purified msVTG was used to produce monoclonal and polyclonal antibodies in mice and rabbits, respectively, and the specificity of the polyclonal antisera for msVTG was confirmed by Western blot analysis. The antibodies cross­reacted with a protein of molecular mass approximately 160 kDa in the plasma samples of mature female marbled sole. No cross-reactivity was observed with the plasma of male fish. A direct non-competitive sandwich enzyme-linked immunosorbent assay (ELISA) was developed using the monoclonal anti-msVTG and polyclonal anti-msVTG antibodies, with purified msVTG as the standard protein. The values of the intra- and inter-assay variations were within the ranges of $8.l-9.8\%$ and $8.5-12.2\%$, respectively. The sensitivity was about 0.3 ng/mL. Serial dilutions of plasma from mature female sole reacted with the msVTG-antibodies in the sandwich ELISA, whereas the plasma from male fish did not. The results indicate that the maturation status of female marbled sole can be identified using a sandwich ELISA for msVTG.

가토에서 방사면역 신티그래피를 이용한 결핵병변의 진단 : 예비보고 (Detection of Tuberculous Lesion by Immunoscintigraphy Using Radiolabeled Specific Polyclonal Antibody Against M. bovis BCG in Rabbit: A Preliminary Result)

  • 이종두;신규호;조상래;신전수;이민걸;양우익;박창윤;유형식;이종태;오옥두;박경배;김재록
    • 대한핵의학회지
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    • 제25권2호
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    • pp.245-251
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    • 1991
  • 결핵성 병변의 단순 x-ray 촬영이나 CT, MRI 소견은 매우 다양하며, 결핵과 전이암 혹은 원발성 암과 감별이 어려운 경우가 있어 결핵으로 확진하기 위하여서 조직 생검이나 수술 등 침습적인 진단 방법을 이용하여야 하였다. 그러므로 이러한 결핵 병변을 비 침습적인 방법으로 정확히 감별할 수 있는 방법을 연구하던 바, 결핵균에 대한 항체를 동위원소에 부착시켜 신티그래피로 진단할 수 있는지의 가능성을 동물실험을 통하여 알아보고자 하였다. 15마리의 가토에서 M.tuberculosis H37Rv를 슬관절에 주입시켜 결핵병변을 유발시키고, 대조군으로 2마리의 가토의 고환에 T.pallidum을 주입하여 매독병변을 유발시킨 후 M.bovis BCG에 대한 특이항체 (specific polyclonal antibody)와 정상 가토의 immunoglobulin을 I-131에 부착시켜 각각의 가토에 주입하여 preset time 10분간 감마카메라로 주사한 결과 다음과 같은 결과를 얻었다. (1) 8마리의 결핵에 감염된 가토에 M.bovis BCG에 대한 $F(ab')_2$를 1 mCi의 I -131 labeling 시킨후 주사한 결과 모두에서 주사후 2시간 부터 72시간까지 병소가 hot uptake으로 보였으며 주사후 24 시간에 가장 높은 target/background ratio를 보였다. (2) 2마리의 매독에 감염된 가토에서 anti-BCG $F(ab')_2$를 주사한 결과 2시간에서는 병소에 hot activity를 보였으나 24시간부터 급격히 activity가 감소하였다. (3) $F(ab')_2$ 대신에 intact antibody를 결핵에 감염된 가토에 투여한 결과 specific polyclonal antibody나 정상가토의 immunoglobulin 모두 결핵병소에 96시간까지 hot uptake를 보였다. 그러므로 결핵균에 대한 specific antibody fragment를 이용하면 방사면역 신티그램으로 진단이 가능하리라 사료되었고, intact antibody를 사용할 경우 sensitivity는 높으나 specificity는 적을 것으로 사료되었다.

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축산식품 중에 잔류하는 Gentamicin 검사를 위한 ELISA 개발에 관한 연구 (Competitive Enzyme-Linked Immunosorbent Assay for Detection of Gentamicin Residues in Edible Animal products)

  • 김재명;이문한;이항;류판동;조명행;박종명
    • 한국식품위생안전성학회지
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    • 제9권3호
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    • pp.123-131
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    • 1994
  • An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)

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