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Recombinat protein containing albumin multimer

A polymer, serum albumin technology, applied in the protein field, can solve the problems of safety, virus contamination, elimination and slow metabolism, and achieve the effect of prolonging the half-life and prolonging the retention time.

Inactive Publication Date: 2005-10-12
NIPRO CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Second, after albumin is released into the blood circulation, its elimination and metabolism are relatively slow
[0005] Meanwhile, pharmaceutical preparations containing albumin from blood can be contaminated with unknown viruses
Therefore, the administration of such pharmaceutical preparations to humans etc. poses a safety problem

Method used

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  • Recombinat protein containing albumin multimer
  • Recombinat protein containing albumin multimer
  • Recombinat protein containing albumin multimer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] What Fig. 1 represented is the general process of embodiment 1 (amplification of human serum albumin gene)

[0042] The human serum albumin gene was integrated into the plasmid pKF18K (hereinafter referred to as pKF18K-HSA, purchased from Tonen General Sekiyu K.K.) (see Fig. 2), and the resulting plasmid was used as a template. With the W-1 sense primer of SEQ.ID.NO.1, the W-1 antisense primer of SEQ.ID.NO.2, the W-2 sense primer of SEQ.ID.NO.3, SEQ.ID. No.4 W-2 antisense primer (as shown in FIG. 8 ) is a synthetic primer, and DNA polymerase (KOD+DNA polymerase, purchased from Toyobo Company) is used for PCR. The reaction conditions of PCR are: process DNA (plasmid) at 94°C for 2 minutes, perform successive denaturation (94°C, 15 seconds), annealing (63°C, 30 seconds) and extension (68°C, 2 minutes) reactions, 30 Cycle, then treat at 68°C for 5 minutes. DNA fragments added with DNA sequences having restriction enzyme sites at the 3' and 5' ends of the DNA sequences en...

Embodiment 2

[0059] Amplification of the DNA sequence between the restriction sites BamHI and XhoI in pPIC9K

[0060] In the same manner as in Example 1, PCR was performed using sense and antisense primers (SEQID. 7 and 8) shown in FIG. 8 and plasmid pPIC9K as a template. The amplified pPIC9K was purified by ethanol precipitation, double-digested with BamHI and XhoI (Takara Shuzo), and the digested fragment was electrophoresed, and then the gel band corresponding to the DNA sequence between BamHI and XhoI was excised . Extracted DNA fragments (2) were obtained from the gel bands using a gel extraction kit (QIAquick Gel Extraction Kit, QIAGEN).

[0061] Ligation and amplification of the human albumin gene

[0062] In the same manner as in Example 1, DNA fragments W-1 (HSA cDNA-1, (3), Fig. 6) and W-2 (HSA cDNA-2, (4), Fig. 6) were obtained from pKF18K-HSA The gel is extracted. The sense and antisense primers (corresponding to SEQ.ID.No.1-4) of DNA fragments W-1 and W-2 are shown in FIG....

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Abstract

A recombinant protein containing a serum albumin multimer wherein at least two genes encoding serum albumin are combined with each other by a gene recombinant technology and allow the production of a serum albumin protein in the host cell is provided. The gene encoding a human serum albumin multimer and the recombinant vector are also provided. The protein is used as a safe drug administration carrier that is capable of extending the half life of a drug in the blood without the risk of contamination of viruses and so on, compared with the conventional drug administration carrier.

Description

Background of the invention [0001] The present invention relates to a protein containing a serum albumin multimer obtained by recombinant technology. More specifically, the present invention relates to a protein, which contains a serum albumin polymer, obtained through recombinant technology, and can be used as a drug delivery carrier, etc., while preventing the risk of infection such as viruses, it can Enhances the retention properties of the drug. Background technique [0002] Human serum albumin (HSA), the major protein found in adult serum, is produced in the liver and functions as a carrier for different serum molecules in humans. In addition, albumin plays an important role in maintaining normal plasma colloid osmotic pressure, which is caused by solutes (colloids) that cannot pass through the pores of capillaries, thereby maintaining the fluid content of the blood. Therefore, albumin has been administered in various surgical treatments, mental injuries, burns, and i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09A61K38/00A61K38/38A61K47/42C07K14/765C12P21/02
CPCA61K38/00C07K14/765
Inventor 小田切优树木田善规片山直久甲斐俊哉
Owner NIPRO CORP
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