Preparation of mouse-human chimeric antibody to an embryonic carbohydrate antigen, Lewis Y.
Stage-specific embryonic antigen-1 (SSEA-1) is a well-known carbohydrate antigen that is specifically expressed on the surface of cancer cells as well as embryonic cells. In this study, starting with a previously established hybridoma producing a monoclonal antibody (named H18A) to Lewis Y antigen, which is closely related to SSEA-1, we cloned genomic DNA encoding active variable regions both of heavy and light chains of the antibody. Sequence analysis showed that VH and V kappa genes of H18A were in the VH 7183 family and V kappa C1 family, respectively. A transfected cell line named HC-H18A-7 expressing a recombinant chimeric H18A composed of mouse-derived antigen-binding variable regions and human-derived constant regions was established. The chimeric H18A was purified to homogeneity and shown to bind purified Lewis Y antigen with the same dose-response curve as the original H18A. The chimeric H18A looks more promising for clinical application than the original mouse-derived H18A because its antigenicity is expected to be reduced.
Kaneko
T
T
Biotechnology Research Laboratory, Tosoh Corporation, Kanagawa.
Iba
Y
Y
Zenita
K
K
Shigeta
K
K
Nakano
K
K
Itoh
W
W
Kurosawa
Y
Y
Kannagi
R
R
Yasukawa
K
K
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D002240:Carbohydrate Sequence; D004247:DNA / Q000737:chemistry; D005803:Genes, Immunoglobulin; D005822:Genetic Vectors; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000235:genetics; D007983:Lewis Blood Group Antigens / Q000737:chemistry / Q000276:immunology*; D016256:Lewis X Antigen / Q000737:chemistry / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D014162:Transfection
D000906:Antibodies; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007983:Lewis Blood Group Antigens; D016256:Lewis X Antigen; D011993:Recombinant Fusion Proteins; D004247:DNA