Methods of producing immunoglobulins, vectors and transformed host cells for use therein
DC CAFC
First Claim
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1. A process for producing an immunoglobulin molecule or an immunologically functional immunoglobulin fragment comprising at least the variable domains of the immunoglobulin heavy and light chains, in a single host cell, comprising the steps of:
- (i) transforming said single host cell with a first DNA sequence encoding at least the variable domain of the immunoglobulin heavy chain and a second DNA sequence encoding at least the variable domain of the immunoglobulin light chain, and (ii) independently expressing said first DNA sequence and said second DNA sequence so that said immunoglobulin heavy and light chains are produced as separate molecules in said transformed single host cell.
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Abstract
The invention relates to processes for producing an immunoglobulin or an immunologically functional immunoglobulin fragment containing at least the variable domains of the immunoglobulin heavy and light chains. The processes can use one or more vectors which produce both the heavy and light chains or fragments thereof in a single cell. The invention also relates to the vectors used to produce the immunoglobulin or fragment, and to cells transformed with the vectors.
755 Citations
36 Claims
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1. A process for producing an immunoglobulin molecule or an immunologically functional immunoglobulin fragment comprising at least the variable domains of the immunoglobulin heavy and light chains, in a single host cell, comprising the steps of:
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(i) transforming said single host cell with a first DNA sequence encoding at least the variable domain of the immunoglobulin heavy chain and a second DNA sequence encoding at least the variable domain of the immunoglobulin light chain, and (ii) independently expressing said first DNA sequence and said second DNA sequence so that said immunoglobulin heavy and light chains are produced as separate molecules in said transformed single host cell. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 19, 34, 35)
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- 15. A vector comprising a first DNA sequence encoding at least a variable domain of an immunoglobulin heavy chain and a second DNA sequence encoding at least a variable domain of an immunoglobulin light chain wherein said first DNA sequence and said second DNA sequence are located in said vector at different insertion sites.
- 18. A transformed host cell comprising at least two vectors, at least one of said vectors comprising a DNA sequence encoding at least a variable domain of an immunoglobulin heavy chain and at least another one of said vectors comprising a DNA sequence encoding at least the variable domain of an immunoglobulin light chain.
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21. A method comprising
a) preparing a DNA sequence consisting essentially of DNA encoding an immunoglobulin consisting of an immunoglobulin heavy chain and light chain or Fab region, said immunoglobulin having specificity for a particular known antigen; -
b) inserting the DNA sequence of step a) into a replicable expression vector operably linked to a suitable promoter;
c) transforming a prokaryotic or eukaryotic microbial host cell culture with the vector of step b);
d) culturing the host cell; and
e) recovering the immunoglobulin from the host cell culture, said immunoglobulin being capable of binding to a known antigen. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32)
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33. A process for producing an immunoglobulin molecule or an immunologically functional immunoglobulin fragment comprising at least the variable domains of the immunoglobulin heavy and light chains, in a single host cell, comprising:
independently expressing a first DNA sequence encoding at least the variable domain of the immunoglobulin heavy chain and a second DNA sequence encoding at least the variable domain of the immunoglobulin light chain so that said immunoglobulin heavy and light chains are produced as separate molecules in said single host cell transformed with said first and second DNA sequences. - View Dependent Claims (36)
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Litigation Data
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Current AssigneeGenentech, Inc. (Roche Holding AG)
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Original AssigneeGenentech, Inc. (Roche Holding AG)
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InventorsCabilly, Shmuel, Wetzel, Ronald B., Riggs, Arthur D., Heyneker, Herbert L., Holmes, William E.
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Primary Examiner(s)Gambel, Phillip
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Application NumberUS07/205,419Time in Patent Office4,939 DaysField of Search435/69.1, 435/69.7, 435/71.1, 435/70.1, 435/71.2, 435/320, 435/261, 435/252.1, 435/252.3, 435/81, 435/55, 435/56, 435/69.6, 435/252.33, 435/254.21, 435/483US Class Current435/69.6CPC Class CodesC07K 16/3007 Carcino-embryonic AntigensC07K 16/462 Igs containing a variable r...C07K 2317/24 containing regions, domains...C07K 2319/00 Fusion polypeptideC07K 2319/02 containing a signal sequenceC07K 2319/30 Non-immunoglobulin-derived ...C12N 15/63 Introduction of foreign gen...C12N 15/81 for yeastsY10S 530/866 involving immunoglobulin or...Y10S 530/867 involving immunoglobulin or...Y10S 930/30 Signal or leader sequence
