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The design, construction and function of a new chimeric anti-CD20 antibody

Paper ID Volume ID Publish Year Pages File Format Full-Text
25374 43570 2007 6 PDF Available
Title
The design, construction and function of a new chimeric anti-CD20 antibody
Abstract

A novel murine IgM-type anti-human CD20 monoclonal antibody (mAb) 1–28 was prepared in our Lab, which can induce apoptosis and inhibit proliferation of Daudi and Raji cells. However, the efficacy of 1–28 mAb in human cancer therapy is likely to be limited by human anti-mouse antibody responses. A chimeric antibody, C1–28, containing 1–28 mAb variable region genes fused to human constant region genes (gamma 1, kappa) was constructed. However, C1–28 lost the antigen-binding activity. Here, using sequence similarity and known 3D structure of antibody variable regions as template, the spatial conformations of 1–28 variable regions (i.e. VH and VL) were analyzed with computer-guided homology modeling methods. According to the surface electrostatic distribution and interaction free energy analysis, the relationship between structure and stability of 1–28 variable regions was studied theoretically and a new chimeric anti-CD20 antibody scFv-Ig named 5S was designed. Expression level of 5S in the culture supernatant was determined to be around 50 μg/mL using sandwich ELISA method with chimeric antibody Rituxan as reference. 5S retained its murine counterpart's binding activity by fluorescence-activated cell-sorting analysis. Furthermore, it could kill CD20 positive Daudi and Raji cells by complement-dependent cytotoxicity. For binding affinity often decreased even lost when IgM antibody was constructed into chimeric IgG1 form, our success give a hint about how to construct a IgG1-type chimeric antibody from IgM-type murine antibody to preserve its binding activity.

Keywords
Computer-guided homology modeling; Chimeric antibody; Complement-dependent cytotoxicity; Single chain Fv
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The design, construction and function of a new chimeric anti-CD20 antibody
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 129, Issue 4, 10 May 2007, Pages 726–731
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us