Construction of single‐chain antibodies that bind an overlapping epitope of HIV‐1 Nef

FEBS Letters

Cassol, Sharon¹  Chang, Alex H²  O'Shaughnessy, Michael²  Hoxie, James A³  Jirik, Frank⁴ 

[1] Division of Infectious Diseases, Department of Medicine, Ottawa General Hospital Research Institute, Box 411, 501 Smyth Rd, Ottawa, Ont. K1H 8L6, Canada;B.C. Center for Excellence in HIV/AIDS, 613–1081 Burrard St., St. Paul's Hospital, Vancouver, B.C. V6Z 1Y6, Canada;Hematology Oncology Section, Rm 664 Clinical Research Building, University of Pennsylvania, 415 Curie Blvd, Philadelphia, PA 19104, USA;Centre for Molecular Medicine and Therapeutics, 950 West 28th Avenue, University of British Columbia, Vancouver, B.C. V5Z 4H4, Canada

关键词: Single-chain antibody;    Monoclonal antibody;    Nef;    Green fluorescent protein;    Human immunodeficiency virus-1;    ScFv;    single-chain Fv (variable fragment) antibody;    Vκ;    variable region of the kappa light chain;    VH;    variable region of the heavy chain;    RT;    reverse transcriptase;    PCR;    polymerase chain reaction;    GFP;    green fluorescent protein;    HIV-1;    human immunodeficiency virus-1;    PBS;    phosphate buffered saline;    GST;    glutathione S-transferase;    CDR;    complementarity determining region;   

DOI  :  10.1016/S0014-5793(98)01569-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The light and heavy chain variable regions of three mouse hybridoma cell lines (AG11, AE6 and EH1) that produce monoclonal antibodies against an overlapping epitope at the C-terminus of Nef were cloned. Sequence analysis of the light and heavy chain variable regions indicated that clones AG11 and AE6, but not EH1, were highly related. Single-chain antibodies were constructed from the cDNA clones of AG11 and EH1, and subcloned into an eukaryotic expressing vector with the green fluorescent protein as marker for expression. Such intracellular antibodies may provide a way in which to inhibit the function of Nef during HIV-1 infection of cells.

【 授权许可】

Unknown