Modular assembly of antibody genes, antibodies prepared thereby and use
DCFirst Claim
1. A process for preparing an immunoglobulin that is capable of binding antigen, said immunoglobulin comprising (a) a heavy chain or heavy chain fragment and (b) a light chain or light chain fragment, said process comprising expressing, in a baceterial cell, of a first nucleic acid that encodes a bacterial signal sequence operably linked to either said heavy chain or to said heavy chain fragment, and a second nucleic acid that encodes a bacterial signal sequence operably linked to either said light chain or to said light chain fragment, such that the bacterial signal sequences bring about the transport of said heavy chain or said heavy chain fragment and said light chain or said light chain fragment through the cytoplasmic membrane of a bacterial cell, and, obtaining said immunoglobulin from the periplasmic space or culture medium, wherein said immunoglobulin is capable of binding antigen.
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Abstract
The invention relates to cDNA genetic sequences, vehicles containing same as well as hosts tansformed therewith, for the production of chimeric immunoglobulin molecules, functional fragments thereof and immunoglobulin derivatives exhibiting novel functional properties comprising human constant region modules and non-human variable region modules, or for class switching antibody molecules and/or chains.
The invention also relates to DNA coding for pectate lyase signal peptide has been cloned on a plasmid to create a secretion vector which is capable of producing a chosen protein which is transported across the bacterial membrane. The secretion vector has been used to secrete extracellular thaumatin and extracellular chimeric antibody fragments. The proteins produced by this vector have biological activity. The thaumatin is properly folded and the antibody fragments are capable of binding antigens on target cancer cells. The invention also relates to the secretion of chimeric antibodies and fragments thereof from yeast in functional form.
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Citations
60 Claims
- 1. A process for preparing an immunoglobulin that is capable of binding antigen, said immunoglobulin comprising (a) a heavy chain or heavy chain fragment and (b) a light chain or light chain fragment, said process comprising expressing, in a baceterial cell, of a first nucleic acid that encodes a bacterial signal sequence operably linked to either said heavy chain or to said heavy chain fragment, and a second nucleic acid that encodes a bacterial signal sequence operably linked to either said light chain or to said light chain fragment, such that the bacterial signal sequences bring about the transport of said heavy chain or said heavy chain fragment and said light chain or said light chain fragment through the cytoplasmic membrane of a bacterial cell, and, obtaining said immunoglobulin from the periplasmic space or culture medium, wherein said immunoglobulin is capable of binding antigen.
- 31. A process for preparing an immunoglobulin that is capable of binding antigen, said immunoglobulin comprising (a) a heavy chain variable region and (b) a light chain variable region, said process comprising operably linking a nucleic acid encoding a bacterial signal sequence to a nucleic acid encoding said heavy chain variable region or said light chain variable region or both of said variable regions such that the bacterial signal sequence or sequences bring about the transport of said variable regions through the cytoplasmic membrane of a bacterial cell, expressing said nucleic acid in said bacterial cell, and obtaining said immunoglobulin from the periplasmic space or culture medium, wherein said immunoglobulin is capable of binding antigen.
Specification