Cj Corporation

Cheil Foods & Chemicals, Inc.

Hyune Soo Lee

Hyun Seok Kim

Myung Suk Oh

Hyung Kon Koh

Yeon Hyang Kim

Jae Hong Lee

Choong S Kim

Ki Wan Kim

Ji Sook Park

Hyung Ki Park

Jong Sang Chung

Yup Kang

Ji W Yoon

Hyune W Lee

Disclosed are human G-CSF isoforms having a modified amino acid sequence containing a glycosylation sequence Asn-X-Ser/Thr (N-X-S/T) at one or more specific amino acid positions according to the present invention, genes encoding the human G-CSF isoforms, and expression vectors carrying the genes, eukaryotic cells transformed or transfected with the expression vectors. Also, the present invention discloses a method of preparing a glycosylated human G-CSF isoform, comprising the steps of culturing the transformant or transfectant and isolating a glycosylated human G-CSF isoform from the culture supernatant or cell lysates, a human G-CSF isoform prepared by the method, and a pharmaceutical composition comprising the human G-CSF isoform.

Glycosylated human granulocyte colony-stimulating factor (G-CSF) isoform

The present invention relates to an amino acid-modified human interferon alpha isoform having at least one of the Asn-Xaa-Ser/Thr (N-X-S/T) sequence formed at a specific site so that glycosylation takes place at this site and a gene encoding the same, an expression vector comprising the gene, and a method for producing glycosylated human interferon alpha isoform by transforming or transfecting an eukaryotic cell with the expression vector, culturing the transfected or transformed cell and isolating the glycosylated human interferon alpha isoform from the culture, the glycosylated human interferon alpha isoform produced therefrom and a pharmaceutical composition comprising the same.

Glycosylated human interferon alpha isoform

The specification describes a process for producing human proinsulin in Escherichia coli (E. coli) using gene manipulation technology. The process can provide for human proinsulin in high yields by a novel expression vector having strong regulatory elements of an insulin gene and a stable recombinant gene product. The expression vector of the present invention is characterized in that: 1) it has an 11 amino acid leader peptide containing six threonines in order to ensure an intracellular stability of proinsulin fusion protein, 2) it contains two copies of a DNA expression cassette each comprising a strong lambda P.sub.R promoter, a lac ribosome binding site, a proinsulin gene with a 17 amino acid leader peptide sequence containing a DNA sequence encoding (Thr).sub.6, and a strong fd phage transcription terminator (combination of phage fd terminator and translation stop codon), etc. successively ligated, 3) it has an ampicillin resistance gene, 4) it can be very stably retained within a cultured cell, and 5) there are a number of these expression vectors in E. coli by which the expression can be significantly increased. Human insulin is prepared from the proinsulin fusion protein by in vitro conversion.

Production of human proinsulin using a novel vector system

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Seoul, South Korea

Hyune Soo Lee