論文 - 詳細
| RRC ID | 44309 |
|---|---|
| 著者 | Han C, Ihara M, Ueda H. |
| タイトル | Expression of an antibody-enzyme complex by the L-chain fusion method. |
| ジャーナル | J Biosci Bioeng |
| Abstract |
In this report, we describe a novel method for directly preparing enzyme-labeled antibodies harvested from IgM-producing hybridoma cells. We constructed expression vectors for antibody light (L) chain-enzyme fusion proteins by linking either the genes for the murine lambda L chain or its constant region (C(L)) with one of two proteins, either the secreted placental alkaline phosphatase or Gaussia luciferase (Gluc). When the vectors were transfected into anti-NP (4-hydroxy-3-nitrophacetyl) IgM-producing myeloma cells, secretion of the IgM-enzyme complex from the gene-transfected cells was confirmed by a direct enzyme-linked immunosorbent assay with an immobilized antigen. Furthermore, when human hybridoma HF10B4, a cell line that produces anti-human lung cancer IgM, was transfected with the vector containing L-Gluc, a significantly stronger signal was obtained for the human lung carcinoma SBC-1 cells than for cervical HeLa cells. Because successful production of an active IgM-enzyme complex containing a heterologous L chain-enzyme fusion was observed, the L-chain fusion method will be a generally applicable method for preparing various IgM-enzyme complexes. |
| 巻・号 | 116(1) |
| ページ | 17-21 |
| 公開日 | 2013-7-1 |
| DOI | 10.1016/j.jbiosc.2013.01.012 |
| PII | S1389-1723(13)00013-3 |
| PMID | 23415663 |
| MeSH | Alkaline Phosphatase / genetics Animals Enzyme-Linked Immunosorbent Assay GPI-Linked Proteins / genetics Genes, Immunoglobulin Light Chain* Genetic Vectors HeLa Cells Humans Hybridomas Immunoglobulin M / genetics Isoenzymes / genetics Luciferases / genetics Lung Neoplasms / genetics Mice Protein Engineering / methods* Recombinant Fusion Proteins / analysis* Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Transfection |
| IF | 2.366 |
| 引用数 | 4 |
| WOS 分野 | FOOD SCIENCE & TECHNOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| リソース情報 | |
| ヒト・動物細胞 | HF10B4(RCB0708) |