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 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.
Sui
Jianhua
J
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
Bai
Jirong
J
St  Clair Tallarico
Aimee
A
Xu
Chen
C
Marasco
Wayne A
WA
D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D015704:CD4 Antigens / Q000276:immunology*; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region; D019169:Jurkat Cells; D008969:Molecular Sequence Data; D019151:Peptide Library; D019718:Receptors, CXCR4 / Q000276:immunology*; D011990:Receptors, Transferrin / Q000276:immunology*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D019718:Receptors, CXCR4; D011990:Receptors, Transferrin