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Modular assembly of antibody genes, antibodies prepared thereby and use
| Details |
Inventors: Robinson, Randy R.; Liu, Alvin Y.; Horwitz, Arnold H.; Better, Marc; Wall, Randolph; Lei, Shau-Ping; Wilcox, Gary L.;
Assignee: Xoma Corporation (Berkeley, CA)
Primary Examiner: Ziska; Suzanne E.
Assistant Examiner:
Attorney, Agent or Firm: Sterne, Kessler, Goldstein, & Fox
The invention relates to cDNA genetic sequences, vehicles containing same as well as hosts transformed 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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DETAILED DESCRIPTION What is claimed is: 1.
A polynucleotide molecule encoding an immunoglobulin fragment, said immunoglobulin fragment comprising a first DNA sequence encoding a first pectate lyase secretion signal sequence operably linked to a DNA sequence encoding an immunoglobulin Fd molecule or a fragment thereof comprising the variable region and a second DNA sequence encoding a second pectate lyase secretion signal sequence operably linked to a DNA sequence encoding an immunoglobulin light chain or a fragment thereof comprising the variable region, wherein said first and second DNA sequences are operably linked to a single prokaryotic promoter so as to form a dicistronic transcription unit, wherein said immunoglobulin fragment is capable of binding to an antigen and said immunoglobulin fragment is produced and secreted by an E.
coli host cell when said polynucleotide molecule is expressed in said host cell.
2.
A vector comprising the polynucleotide molecule of claim 1.
3.
An E.
coli host cell transformed with the vector of claim 2.
4.
The polynucleotide molecule of claim 1, wherein said immunoglobulin Fd molecule or said immunoglobulin light chain are selected such that the resultant immunoglobulin fragment is chimeric.
Description:
FIELD OF THE INVENTION This invention relates to recombinant DNA methods of preparing immunoglobulins, genetic sequences coding therefor, as well as methods of obtaining such sequences.
BACKGROUND ART The application of cell-to-cell fusion for the production of monoclonal antibodies by Kohler and Milstein (Nature (London), 256: 495, 1975) has spawned a revolution in biology equal in impact to the invention of recombinant DNA cloning.
Hybridoma-produced monoclonal antibodies are already widely used in clinical diagnoses and basic scientific studies.
Applications of human B cell hybridoma-produced monoclonal antibodies hold great promise for the clinical treatment of cancer, viral and microbial infections, B cell immunodeficiencies with diminished antibody production, and other diseases and disorders of the immune system
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