Humanization and pharmacokinetics of a monoclonal antibody with specificity for both E- and P-selectin
| Autor: | Xy, He, Xu Z, Melrose J, Mullowney A, Vasquez M, Queen C, Vexler V, Klingbeil C, Ms, Co, Ellen Berg |
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| Rok vydání: | 1998 |
| Předmět: |
Models
Molecular Mice Inbred BALB C DNA Complementary Base Sequence Recombinant Fusion Proteins Immunology Molecular Sequence Data Antibody Affinity Immunoglobulin Variable Region Antibodies Monoclonal Binding Competitive Macaca mulatta Mice P-Selectin Antibody Specificity Cell Adhesion Immunology and Allergy Animals Humans Amino Acid Sequence Cloning Molecular E-Selectin Half-Life |
| Zdroj: | Europe PubMed Central |
| ISSN: | 0022-1767 |
| Popis: | E- and P-selectin (CD62E and CD62P) are cell adhesion molecules that mediate leukocyte-endothelial cell and leukocyte-platelet interactions and are involved in leukocyte recruitment during inflammation. We previously developed a murine mAb, EP-5C7 (or mEP-5C7), that binds and blocks both E- and P-selectin. When used in humans, murine mAbs have short circulating half-lives and generally induce potent human anti-mouse Ab responses. We therefore engineered a humanized, complementarity determining region-grafted version of mEP-5C7 incorporating human γ4 heavy and κ light chain constant regions (HuEP5C7.g4). HuEP5C7.g4 retains the specificity and avidity of mEP-5C7, binding to human E- and P-selectin but not to human L-selectin, and blocking E- and P-selectin-mediated adhesion. Surprisingly, when administered to rhesus monkeys, HuEP5C7.g4 was eliminated from the circulation very rapidly, even faster than the original murine Ab. To isolate the cause of the short serum half-life of HuEP5C7.g4, several Ab variants were constructed. A chimeric IgG4 Ab was made by replacing the humanized V regions with murine V regions. A humanized IgG2 Ab, HuEP5C7.g2, was also made by replacing the human γ4 with a γ2 constant region. Results from pharmacokinetic studies in rhesus monkeys demonstrated that the chimeric IgG4 is also rapidly eliminated rapidly from serum, similar to the humanized IgG4 Ab, while the humanized IgG2 Ab displays a long circulation half-life, typical of human Abs. |
| Databáze: | OpenAIRE |
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