Polymerase chain reaction facilitates the cloning, CDR-grafting, and rapid expression of a murine monoclonal antibody directed against the CD18 component of leukocyte integrins.
نویسندگان
چکیده
Two novel approaches of recombinant PCR technology were employed to graft the complementarity determining regions from a murine monoclonal antibody (mAb) onto human antibody frameworks. One approach relied on the availability of cloned human variable region templates, whereas the other strategy was dependent only on human variable region protein sequence data. The transient expression of recombinant humanized antibody was driven by the adenovirus major late promoter and was detected 48 hrs post-transfection into non-lymphoid mammalian cells. The application of these new approaches enables the expression of a recombinant humanized antibody just 6 weeks after initiating the cDNA cloning of the murine mAb.
منابع مشابه
THE PRODUCTION OF MURINE MONOCLONAL ANTIBODIES (MAb) DIRECTED AGAINST HUMAN T- LYMPHOCYTE SUBSETS
The production of murine monoclonal antibody (MAb) has not yet been reported in Iran. The present work describes for the first time the generation of several murine hybridoma clones secreting MAbs directed against human leukocyte surface antigens. The secreted antibodies by hybridoma clones have been screened on different lymphoid and non-lymphoid tissues. Results indicated that of seven h...
متن کاملDesign and Construction of a Novel Humanized Single-Chain Variable-Fragment Antibody against the Tumor Necrosis Factor alpha
The pro-inflammatory cytokine, TNF-α, which plays a major role in the development and persistence of inflammatory diseases, is the basis for the use of anti-TNF-α therapies. The neutralization of TNF-α or blockage of its binding to the corresponding receptor has mainly served as a therapeutic strategy against some diseases. This study aimed to investigate the production of a humanized single ch...
متن کاملDesign and Construction of a Novel Humanized Single-Chain Variable-Fragment Antibody against the Tumor Necrosis Factor alpha
The pro-inflammatory cytokine, TNF-α, which plays a major role in the development and persistence of inflammatory diseases, is the basis for the use of anti-TNF-α therapies. The neutralization of TNF-α or blockage of its binding to the corresponding receptor has mainly served as a therapeutic strategy against some diseases. This study aimed to investigate the production of a humanized single ch...
متن کاملتولید آنتی بادی تک زنجیرهای انسانی شده ضد مارکر CD20 در E.coli
Background and Objectives: Rituximab is an anti-CD20 chimeric monoclonal antibody widely used for the treatment of malignant B cells lymphoma. However, the immunogenicity of murine-derived monoclonal antibodies and the large size of full length antibodies restrict cancer immunotherapy. Humanized single chain antibodies can be a solution and a promising alternative for application in immunothera...
متن کاملCloning and secretory expression of VP2 gene of infectious bursal disease virus in eukaryotic cells
VP2 gene coding region of a vaccinal strain (D78) of infectious bursal disease virus (IBDV) was clonedin a eukaryotic expression vector, pSec Tag2A. The gene was placed downstream of Ig κ chain leadersequence, under the control of human cytomegalovirus (hCMV) immediate early enhancer and promoter. Theconstruct pSec Tag2A-VP2 was transfected in COS-7 cell line and the expression and secretion of...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Nucleic acids research
دوره 19 9 شماره
صفحات -
تاریخ انتشار 1991