The United States of America as Represented by the Department of Health

The United States of America, as Represented by the Secretary, Department of Health and Human Services

Sun-Hee Jung

Ira H Pastan

Byungkook Lee

Ulrich Brinkmann

Eduardo Agustin Padlan

Itai Benhar

The present invention relates to disulfide-stabilized recombinant polypeptide molecules which have the binding ability and specificity for another peptide, such as the variable region of an antibody molecule. Methods of producing these molecules and nucleic acid sequences encoding these molecules are also described. In particular, the invention discloses Fv antibody fragments stabilized by a disulfide bond connecting the V and V regions of the Fv fragment. The &agr; and &bgr; chains of T cell receptors may be similarly stabilized by means described in the invention.

Methods of making recombinant disulfide-stabilized polypeptide fragments having binding specificity

The invention provides methods for inhibiting the growth of a cell bearing a Lewis,antigen. The methods involve contacting the cell with a composition comprising an Fv region of a light chain of a monoclonal antibody selected from B1, B3, and B5, and the Fv region of a humanized heavy chain of a monoclonal antibody independently selected from B1, B3, and B5, provided that if the heavy and the light chains are from the same antibody, the residue at position 95 of the heavy chain is a serine, and, if the antibody chain is from B3, the residue at position 4 of the light chain can be a leucine and the residue at position 82,of the heavy chain can be an arginine. The Fv regions are joined to an effector molecule selected from a chemotherapeutic agent, a toxin, a radioisotope, and a liposome loaded with a chemotherapeutic agent. In preferred embodiments, the heavy chain and the light chain are recombinantly fused, and the Fv regions are recombinantly fused to a toxin.

Methods of inhibiting the growth of cells bearing LewisY antigens using B1, B3, or B5 targeted immunoconjugates

The present invention relates to disulfide-stabilized recombinant polypeptide molecules which have the binding ability and specificity for another peptide, such as the variable region of an antibody molecule. Methods of producing these molecules and nucleic acid sequences encoding these molecules are also described. In particular, the invention discloses Fv antibody fragments stabilized by a disulfide bond connecting the V.sub.H and V.sub.L regions of the Fv fragment. The .alpha. and .beta. chains of T cell receptors may be similarly stabilized by means described in the invention.

Recombinant disulfide-stabilized polypeptide fragments having binding

This invention provides for recombinant single chain antibodies capable of specifically binding to a Lewis.sup.Y -related carbohydrate antigen and fusion proteins comprising these antibodies. More particularly, the invention provides for humanized chain Fv regions of the monoclonal antibodies B1, B3 and B5 and fusion proteins incorporating these humanized antibodies. The antibodies may comprise humanized variable heavy (V.sub.H) chains, humanized variable light (V.sub.L) chains, or both. The invention also provides for DNA sequences encoding the various humanized antibodies. In addition, the invention provides for methods of detecting cells bearing a Lewis.sup.Y antigen in a patient and for methods of killing or inhibiting the growth of cells bearing a Lewis.sup.Y antigen in a patient.

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Bethesda, MD, United States of America

Sun-Hee Jung