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한국학술지인용색인(NRF)
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Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FITC conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt`s lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt`s lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.
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참고문헌 (35건) : 자료제공( 네이버학술정보 )
| 번호 | 참고문헌 | 국회도서관 소장유무 |
|---|---|---|
| 1 | Taking Aim at Cancer with Monoclonal Antibodies  |
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| 2 | Clark TJ. L'impact économique des anticorps monoclonaux. Biofutur 1986;44:121-33. | 미소장 |
| 3 | Production of the Recombinant Single Chain Anti-B Cell Lymphoma Antibody and Evaluation of Immunoreactivity | 소장 |
| 4 | Enhancement of the activity of immunotoxins made with either ricin A chain or Pseudomonas exotoxin in human ovarian and epidermoid carcinoma cell lines. |
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| 5 | Radioimmunotherapy of human colonic cancer xenografts with 90Y labeled monoclonal antibodies to carcinoembryonic antigen. |
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| 6 | ANTIBODY-GUIDED IRRADIATION OF MALIGNANT LESIONS: THREE CASES ILLUSTRATING A NEW METHOD OF TREATMENT: A Report from the Hammersmith Oncology Group and the Imperial Cancer Research Fund |
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| 7 | Use of isotopic immunoglobulin in therapy. |
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| 8 | Molecular cloning of a cDNA encoding a bovine growth differentiation factor-9 (GDF-9) and expression of GDF-9 in bovine ovarian oocytes and in vitro-produced embryos. |
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| 9 | Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase |
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| 10 | Deciphering the Mechanism for the Assembly of Aromatic Polyketides by a Bacterial Polyketide Synthase |
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| 11 | Optimization of an Osmotic Shock Procedure for Isolation of a Protein Product Expressed in E. coli |
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| 12 | Jutta Drees, Jason Smith, Frank Schäfer, Kerstin Steinert. The QIA expressionist a handbook for high-level expression and purification of 6X His-tagged proteins. Germany: QIAGEN GmbH, Hilden; 2002. | 미소장 |
| 13 | Coligan JE, Dunn BM, Ploegh HL, Speicher DW, Wingfield PT. Current protocols in protein science. New York: John Wiley and Sons; 1995. | 미소장 |
| 14 | Rapid and Efficient Purification of Native Histidine-Tagged Protein Expressed by Recombinant Vaccinia Virus |
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| 15 | Darby N, Creighton TE. In Protein Stability and folding: Theory and Practice. USA: Humana Press, Inc; 1995. | 미소장 |
| 16 | Ausubel FM, Brent R, Kingston RE. Current protocols in molecular biology Vol. 1. New York: John Wiley and Sons; 1994. | 미소장 |
| 17 | Methods in Enzymology |
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| 18 | Maniatas T, Frisch EF, Sambrook MD. Molecular cloning: A laboratory manual. New York: Cold Spring Harbor Laboratory; 1989. | 미소장 |
| 19 | Cleavage of structural proteins during the assembly of the head of bacteriophage T4. |
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| 20 | Reduction-mediated technetium-99m labeling of monoclonal antibodies. |
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| 21 | Hong MK, Jeong JM, Yeo JS, Kim KM, Chang YS, Lee YS, et al. In vitro properties and biodistribution of Tc-99m and Re-188 labeled monoclonal antibody CEA 79.2. Korean J Nucl Med 1988; 32:516-24. | 미소장 |
| 22 | Preparation and comparative evaluation of 99mTc-labeled 2-iminothiolane modified antibodies and CITC-DTPA immunoconjugates of anti-EGF-receptor antibodies. |
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| 23 | Egan JA, Filer CN. Tritium labeling of the local anesthetic tetracaine via a polyhalogenated precursor. J Radioanal Nucl Chem 2003;10:1023-9. | 미소장 |
| 24 | The B cell surface molecule B1 is functionally linked with B cell activation and differentiation. |
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| 25 | Expression of human B cell-associated antigens on leukemias and lymphomas: a model of human B cell differentiation. |
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| 26 | Retention of B-cell-specific monoclonal antibodies by human lymphoma cells. |
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| 27 | Binding to CD20 by anti-B1 antibody or F(ab')(2) is sufficient for induction of apoptosis in B-cell lines. |
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| 28 | The application of fluorescein isothiocyanate and high-performance liquid chromatography for the microsequencing of proteins and peptides |
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| 29 | Highly fluorochrome labeled gene probes for quantitative tracing of RNA in individual cells by in situ hybridization |
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| 30 | ZAP-70 expression identifies a chronic lymphocytic leukemia subtype with unmutated immunoglobulin genes, inferior clinical outcome, and distinct gene expression profile. |
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| 31 | Expression of ZAP-70 is associated with increased B-cell receptor signaling in chronic lymphocytic leukemia. |
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| 32 | Flow cytometric analysis of cell-surface and intracellular antigens in leukemia diagnosis. |
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| 33 | Protein aggregation as bacterial inclusion bodies is reversible |
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| 34 | Construction and deconstruction of bacterial inclusion bodies |
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| 35 | Western blot analysis of antibody response to pneumococcal protein antigens in a murine model of pneumonia. |
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