TAILIEUCHUNG - Báo cáo khoa học: Identification of CD4 and transferrin receptor antibodies by CXCR4 antibody-guided Pathfinder selection

To generate human antibodies against CXCR4, a seven-transmembrane chemokine receptor and a principal coreceptor forHIV-1, several roundsofPathfinder andStep-back selection from a large phage display antibody library were performed on Jurkat cells. A mAb against CXCR4 or biotinyated phage antibodies were used as guide molecules. Over 100 pan-Jurkat-cell-positive antibodies were charac-terized, but none were CXCR4 specific. | Eur. J. Biochem. 270 4497-4506 2003 FEBS 2003 doi Identification of CD4 and transferrin receptor antibodies by CXCR4 antibody-guided Pathfinder selection Jianhua Sui1 2 Jirong Bai1 Aimee St. Clair Tallarico1 Chen Xu1 2 and Wayne A. Marasco1 2 1Department of Cancer Immunology AIDS Dana-Farber Cancer Institute Boston MA 02115 USA department of Medicine Harvard Medical School Boston MA 02115 USA To generate human antibodies against CXCR4 a seven-transmembrane chemokine receptor and a principal coreceptor for HIV-1 several rounds of Pathfinder and Step-back selection from a large phage display antibody library were performed on Jurkat cells. A mAb against CXCR4 or biotinyated phage antibodies were used as guide molecules. Over 100 pan-Jurkat-cell-positive antibodies were characterized but none were CXCR4 specific. However several antibodies against CD4 and the transferrin receptor were identified. Our results indicate that although Pathfinder and Step-back selection can be used to select phage antibodies on whole cells the successful selection of certain targets is still complex and limited. The reason is probably in part due to the inaccessibility of the targeted extracellular structures and the range of the horseradish peroxidase-labeled guide molecule. Refinements of these techniques are required to improve target specificity and selectivity. Keywords CXCR4 Pathfinder selection phage display scFv transferrin receptor. Naive and nonimmune antibody phage libraries are a powerful tool that enables the selection and identification of antibodies to purified antigens 1-3 . In some cases in which potential therapeutic or research antibodies against cell surface antigens are required phage screening is performed on whole cells or cell membranes. However cell based screening is often difficult because of the much greater antigen complexity lower antigen concentration and antigen inaccessibility. Some selection methods using whole cells have .

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