Humanization by variable domain resurfacing and grafting on a human IgG4, using a new approach for determination of non-human like surface accessible framework residues based on homology modelling of variable domains.

摘要: Abstract Many antithrombotic agents have only a small therapeutic window, frequently leading to bleeding problems. However, interfering with platelet adhesion through the collagen-VWF-GPIbα axis is expected cause less Our group developed monoclonal antibody, 82D6A3, directed against von Willebrand factor (VWF) A3-domain, which inhibits VWF-interaction fibrillar collagen. 82D6A3 has effects in vivo without time prolongation. To further investigate promising features of we humanized it by variable domain resurfacing and grafting on constant regions human IgG 4 . First, sequence domains was determined murine scFv constructed. The expressed had comparable activity as its DNA thus used subsequent humanization procedures. For this, new approach introduced identify non-human like framework surface residues, since general distribution accessible residues described for heavy light chain showed several discrepancies homology modelled Fv 82D6A3. Identification evaluation their accessibility within context revealed 10 that need be influencing conformation CDR loops. Indeed, obtained mutating all counterpart, still binding high affinity VWF retained inhibitory properties scFv. Next, order increase half life decrease immunogenicity, were grafted , resulting h82D6A3 an vitro IgG.