Humanization of high affinity anti-HBs antibody by using human consensus sequence and modification of selected minimal positional template and packing residues.
We had earlier reported the construction and characterization of a high affinity recombinant scFv generated from a potential neutralizing mouse monoclonal antibody against the Hepatitis B surface antigen. In this report we describe the humanization of this scFv by grafting its antigen binding site onto framework of the human consensus sequence of highest similarity. We have used molecular modeling to alter not only the clearly permissible residues but also several minimal positional template and V(H)/V(L) interface residues. The humanized scFv retains the binding characteristic of the mouse monoclonal even under conditions that usually destabilize antigen antibody interactions. This high affinity humanized scFv provides a basis for the development of prophylactic/therapeutic molecules.
Tiwari
Ashutosh
A
Department of Biochemistry, All India Institute of Medial Sciences, New Delhi, 110029, India. ashutosht@hotmail.com
Khanna
Navin
N
Acharya
Subrat K
SK
Sinha
Subrata
S
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D000915:Antibody Affinity; D000918:Antibody Specificity; D016384:Consensus Sequence / Q000276:immunology*; D006510:Hepatitis B Antibodies / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology*; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D007158:Immunologic Techniques; D051379:Mice; D018448:Models, Immunological; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis
D000911:Antibodies, Monoclonal; D006510:Hepatitis B Antibodies; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins