Baylor College of Medicine

Agennix Incorporated

Other

Orla M Conneely

Bert W O'Malley

Denis R Headon

Gregory S May

Ronan F Power

Pauline P Ward

Production of recombinant lactoferrin and lactoferrin polypeptides using cDNA sequences in various organisms

The present invention is directed to recombinant nucleic acids encoding lactoferrin variants and portions thereof, having modified iron-binding capacity, and to vectors comprising same recombinant nucleic acids. The present invention is further directed to methods of producing such vectors, and to transfected cells harboring the same. Methods for the production of lactoferrin variants and portions thereof, in various eukaryotic or prokaryotic cells are also provided. Finally, the invention is directed to lactoferrin variants and portions thereof encoded by the nucleic acids of the invention and produced by the processes of the invention. Thus, the invention provides an efficient and economical means for the production of recombinant lactoferrin variants and portions thereof.

Lactoferrin variants and uses thereof

The verified cDNA sequences for human, bovine and porcine lactoferrin protein have been used to prepare recombinant lactoferrin for therapeutic and nutritional applications. Regions of the cDNA such as the Fe binding sites can be used to make an hLF polypeptide product. The present invention provides novel plasmids, transfected eucaryotic cells and methods of producing these plasmids and transfected eucaryotic cells. The novel plasmid contains the cDNA for lactoferrin protein. Methods for the production of lactoferrin protein in fungi and bacteria are also provided. Thus, the present invention provides an efficient and economical means for the production of recombinant lactoferrin protein and lactoferrin related polypeptides.

Production for recombinant lactoferrin and lactoferrin polypeptides

The subject invention provides for the production of lactoferrins and lactoferrin polypeptide fragments using the host cells Aspergillus in combination with novel plasmid constructs. More specifically, the subject invention provides novel vector constructs capable of producing lactoferrins and lactoferrin polypeptide fragments in Aspergillus host cells. More particularly, the subject invention provides for novel plasmid constructs suitable for use with Aspergillus and especially Aspergillus awamori, niger and oryzae host cells, which enables them to produce large amounts of recombinant lactoferrins and lactoferrin polypeptide fragments.

Expression of processed recombinant lactoferrin and lactoferrin

A tissue culture screening system to monitor a transcriptional response treated by a chemical signal interacting with a plasma membrane receptor is provided. The tissue culture screening system includes a cell line containing a membrane receptor, a target gene and a specific receptor selected from the group consisting of a steroid receptor, a vitamin receptor and an orphan receptor. The specific receptor regulates transcription of the target gene. Any of the target gene membrane receptor or specific receptor can be introduced into the cell by an expression vector. In addition to the screening system there is also provided assays for identifying test compounds and chemical signals that regulate transcription or are potential agonist or antagonist neurotransmitters or which regulate indirectly by a membrane receptor binding or regulate transription in the absence of a steroid, vitamin or orphan ligand. There is further provided kits for the assays.

Screening system for identifying compounds that regulate steroid and

Production of recombinant lactoferrin and lactoferrin polypeptides using

A tissue culture screening system to monitor a transcriptional response treated by a chemical signal interacting with a plasma membrane receptor is provided. The tissue culture screening system includes a cell line containing a membrane receptor, a target gene and a specific receptor selected from the group consisting of a steroid receptor, a vitamin receptor and an orphan receptor. The specific receptor regulates transcription of the target gene. Any of the target gene membrane receptor or specific receptor can be introduced into the cell by an expression vector. In addition to the screening system there is also provided assays for identifying test compounds and chemical signals that regulate transcription or are potential agonist or antagonist neurotransmitters or which regulate indirectly by a membrane receptor binding or regulate transcription in the absence of a steroid, vitamin or orphan ligand. There is further provided kits for the assays.

Screening system and assay for identifying compounds that regulate

The present invention provides novel plasmids, transfected eucaryotic cells and methods of producing these plasmids and transfected eucaryotic cells. The novel plasmid contains the cDNA for human lactoferrin protein. Methods for the production of human lactoferrin protein in A. Oryzae are also provided. Thus, the present invention provides an efficient and economical means for the production of recombinant human lactoferrin protein.

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Houston, TX, United States of America

Orla M Conneely