Anti-idiotypic antibody as the surrogate antigen for cloning scFv and its fusion proteins Journal Article
| Authors: | Cheung, N. K. V.; Guo, H. F. ; Modak, S.; Cheung, I. Y. |
| Article Title: | Anti-idiotypic antibody as the surrogate antigen for cloning scFv and its fusion proteins |
| Abstract: | Single-chain variable fragment (ScFv) is a versatile building block for novel targeting constructs. However, a reliable screening and binding assay is often the limiting step for antigens that are difficult to clone or purify. Anti-idiotypic antibodies may be useful as surrogate antigens for cloning scFv and their fusion proteins. 8H9 is a murine IgG1 monoclonal antibody (MAb) specific for a novel antigen expressed on the cell surface of a wide spectrum of human solid tumors, but not in normal tissues. Rat anti-SH9-idiotypic hybridomas (clones 2E9, 1E12, and 1F11) were produced by somatic cell fusion between rat lymphocytes and mouse SP2/0 myeloma. In direct binding assays enzyme-linked immunosorbant assay - (ELISA) - they were specific for the 8H9 idiotope. Using 2E9 as the surrogate antigen, 8Hg-scFv was cloned from hybridoma cDNA by phage display. 8H9scFv was then fused to human-γ1-CH2-CH3 cDNA for transduction into CHO and NSO cells. High expressors of mouse scFv-human Fc chimeric antibody were selected. The secreted homodimer reacted specifically with antigen-positive tumor cells by ELISA and by flow cytometry, inhibitable by the anti-idiotypic antibody. The reduced size resulted in a shorter half-life in vivo, while achieving comparable tumor to nontumor ratio as the native antibody 8H9. However, its in vitro activity in antibody-dependent cell-mediated cytotoxicity was modest. |
| Keywords: | signal transduction; controlled study; human cell; nonhuman; flow cytometry; animal cell; mouse; animals; mice; animal tissue; animal experiment; animal model; enzyme linked immunosorbent assay; mice, inbred balb c; animalia; monoclonal antibody; molecular cloning; cloning, molecular; immunoglobulin variable region; antigen; hybrid protein; recombinant fusion proteins; rat; murinae; antibody specificity; lymphocyte; single chain fragment variable antibody; phage display; antibody production; complementary dna; antibody dependent cellular cytotoxicity; cell fusion; chimeric antibody; myeloma cell; antibodies, anti-idiotypic; antiidiotypic antibody; human; priority journal; article |
| Journal Title: | Hybridoma and Hybridomics |
| Volume: | 21 |
| Issue: | 6 |
| ISSN: | 1536-8599 |
| Publisher: | Mary Ann Liebert, Inc |
| Date Published: | 2002-12-01 |
| Start Page: | 433 |
| End Page: | 443 |
| Language: | English |
| PUBMED: | 12573107 |
| PROVIDER: | scopus |
| DOI: | 10.1089/153685902321043963 |
| DOI/URL: | |
| Notes: | Export Date: 14 November 2014 -- Source: Scopus |
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