EP0173494A2
Expired / Lapsed
EP
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Chimeric receptors by DNA splicing and expression
Assignee: Leland Stanford Junior University
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Filing: 1985-08-07
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Granted: —
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Expires: 2005-08-07
Abstract
Abstract Chimeric multi-subunit receptors are provided involving a binding region resulting from the organization of two subunit chains and a non-binding region which may have physiological significance. Particularly, chimeric immunoglobulins are provided having variable regions from one species and constant regions from another species, by linking DNA sequences encoding for the variable regions of the light and heavy chains from one species to the constant regions of the light and heavy chains respectively from a different species. Introduction of the resulting genes into mammalian host cells under conditions for expressions provides for production of chimeric immunoglobulins having the specificity of the variable region derived from a first species and the physiological functions of the constant region from a different species.
Inventors
- 👤 Sherie L. Morrison
- 👤 Leonard A. Herzenberg
- 👤 Vernon T. Oi
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 17:53
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/46; C07K16/461; C07K16/462; A; A61; A61K; A61K38/00; C07K2317/00; C07K2317/20; C07K2317/24; C07K2319/00; C07K2319/01; C07K2319/02
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
121 words
A multi-subunit chimeric receptor with at least one binding site and having two subunits which are bound together and define a predetermined ligand binding site, wherein each of said two subunits consists of a variable region which participates in defining said binding site and binding to said ligand and a non-binding region (constant region) which participates in maintaining the structure of said receptor, wherein each of said variable regions has an amino acid sequence having substantially the same amino acid sequence of a variable region of a receptor from a first mammalian source and each of said constant regions has an amino acid sequence having substantially the same amino acid sequence of a constant region from a different second mammalian source.
Claim 2
Dependent Claim
121 words
A multi-subunit chimeric receptor with at least one binding site and having two subunits which are bound together and define a predetermined ligand binding site, wherein each of said two subunits consists of a variable region which participates in defining said binding site and binding to said ligand and a non-binding region (constant region) which participates in maintaining the structure of said receptor, wherein each of said variable regions has an amino acid sequence having substantially the same amino acid sequence of a variable region of a receptor from a first mammalian source and each of said constant regions has an amino acid sequence having substantially the same amino acid sequence of a constant region from a different second mammalian source.
Claim 3
Dependent Claim
14 words
A multi-subunit chimeric receptor according to Claim 1, wherein said receptor is an immunoglobulin.
Claim 4
Dependent Claim
19 words
A multi-subunit chimeric receptor according to Claim 1 or Claim 2, joined to a label providing a detectable signal.
Claim 5
Dependent Claim
93 words
A DNA construct for expression of a chimeric polypeptide which is a subunit of a multi-unit receptor comprising: a 5'-terminus proximal first DNA sequence coding for a polypeptide substantially the same as the N-terminal variable binding region of a receptor from a first mammalian host; and a 3'-terminus proximal second DNA sequence joined directly or through an intron in reading frame at its 5' end to the 3' end of said first DNA sequence, coding for a polypeptide substantially the same as the non-binding region (constant region) from a different second mammalian host.
Claim 6
Dependent Claim
93 words
A DNA construct for expression of a chimeric polypeptide which is a subunit of a multi-unit receptor comprising: a 5'-terminus proximal first DNA sequence coding for a polypeptide substantially the same as the N-terminal variable binding region of a receptor from a first mammalian host; and a 3'-terminus proximal second DNA sequence joined directly or through an intron in reading frame at its 5' end to the 3' end of said first DNA sequence, coding for a polypeptide substantially the same as the non-binding region (constant region) from a different second mammalian host.
Claim 7
Dependent Claim
29 words
A DNA construct according to Claim 4, having transcriptional initiation and termination regulatory sequences recognized by a mammalian host in regulatory relationship with said first and second DNA sequences.
Claim 8
Dependent Claim
23 words
A DNA construct according to Claim 4 or Claim 5, wherein said first and second DNA sequences code for an immunoglobulin light chain.
Claim 9
Dependent Claim
23 words
A DNA construct according to Claim 4 or Claim 5, wherein said first and second DNA sequences code for an immunoglobulin heavy chain.
Claim 10
Dependent Claim
145 words
A mammalian cell having first and second DNA constructs for expression of different first and second subunits of a multi-unit receptor, said first and second subunits when bound together defining a binding site for a predetermined ligand; each of said first and second constructs comprising: a 5'-terminus proximal first DNA sequence coding for a polypeptide substantially the same as the N-terminal variable binding region of a receptor from a first mammalian host; and a 3'-terminus proximal second DNA sequence joined directly or through an intron in reading frame at its 5' end to the 3' end of said first DNA sequence coding for a polypeptide substantially the same as the non-binding region (constant region) from a different second mammalian host; and transcriptional and translational regulatory initiation and termination sequences functional in said mammalian cell and in regulatory relationship with said first and second DNA constructs.
Claim 11
Dependent Claim
145 words
A mammalian cell having first and second DNA constructs for expression of different first and second subunits of a multi-unit receptor, said first and second subunits when bound together defining a binding site for a predetermined ligand; each of said first and second constructs comprising: a 5'-terminus proximal first DNA sequence coding for a polypeptide substantially the same as the N-terminal variable binding region of a receptor from a first mammalian host; and a 3'-terminus proximal second DNA sequence joined directly or through an intron in reading frame at its 5' end to the 3' end of said first DNA sequence coding for a polypeptide substantially the same as the non-binding region (constant region) from a different second mammalian host; and transcriptional and translational regulatory initiation and termination sequences functional in said mammalian cell and in regulatory relationship with said first and second DNA constructs.
Claim 12
Dependent Claim
27 words
A mammalian cell according to Claim 8, wherein said first DNA construct encodes an immunoglobulin light chain and second second DNA construct encodes an immunoglobulin heavy chain.
Claim 13
Dependent Claim
44 words
A method for producing a chimeric receptor which comprises: growing mammalian cells according to Claim 8 or Claim 9 in a nutrient medium, whereby said DNA constructs are expressed and said subunits are intracellularly bound together to form a receptor; and isolating said receptor.
Claim 14
Dependent Claim
14 words
A method according to Claim 10, wherein said cell is a mouse myeloma cell.
Claim 15
Dependent Claim
27 words
A method according to Claim 11, wherein said mouse myeloma cell is grown in the peritoneal cavity of a mouse resulting in the production of ascites fluid.
References Cited (Backward Citations)
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