Establishment, molecular rescue, and expression of 123AV16-1, a tumor-reactive human monoclonal antibody.
The human monoclonal antibody (mAb) 123AV16-1 was generated by Epstein-Barr virus transformation of peripheral blood lymphocytes from a colorectal patient undergoing active specific immunotherapy with an autologous tumor cell-Bacille Calmette-Guérin vaccine. Direct immunohistochemical staining of tumor and normal pairs of tissues indicated that this human IgA1, lambda 2 mAb preferentially reacted with colon tumor epithelium. To generate a recombinant derivative of this Epstein-Barr virus-transformed cell line, we isolated the expressed complete heavy and light chain genes by a novel strategy and cloned them into modified pSV2-neo and pSV2-gpt expression vectors. The recombinant 123AV16-1 human mAb was expressed in both a murine myeloma and a human-murine heteromyeloma and was secreted as both monomers and dimers. The recombinant 123AV16-1 mAb expressed by both cell lines reacted with human colon tumor xenografts in a manner similar to the mAb derived from the Epstein-Barr virus-transformed cell line, indicating that the antibody specificity was not appreciably altered during the molecular rescue, cloning, or expression.
Hall
B L
BL
Organon Teknika Biotechnology Research Institute, Rockville, Maryland 20850.
Murray
J H
JH
Haspel
M V
MV
Kobrin
B J
BJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D001483:Base Sequence; D002461:Cell Line, Transformed; D015179:Colorectal Neoplasms / Q000276:immunology*; D005260:Female; D004854:Herpesvirus 4, Human; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis*
D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins