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Recombinant fusion protein with targeted cell for killing tumor

A technology of fusion protein and Pseudomonas, which is applied in the field of recombinant fusion protein with the ability to kill tumor cells, can solve the problems of high dosage and low biological activity, and achieve the effect of improving activity

Inactive Publication Date: 2006-10-04
BEIJING NORTHLAND BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology describes an approach called targeting fused protein(TFP). It can be applied effectively against certain types of diseases like lung cancer or melanoma by delivering it into specific areas where they will attack them more efficiently than other treatments. These TFPs were found to improve their ability to kill these targets while also being less harmful when delivered through injection methods.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the delivery of drugs specifically towards targets like cancer tissue without causing harmful healthy ones. Current methods are either expensive or result in poor bioactivity due to their lack of selective engagement ability against other parts of the body's own cells.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0009] According to the protein sequence of fusion protein 1 and the principle of codon preference of hosts in different expression systems, the prokaryotic expression gene sequence and eukaryotic expression gene sequence of fusion protein 1 were designed, and synthesized by the method of whole gene synthesis combined with PCR. After the gene sequence is verified by sequencing, the target gene is connected into the expression vector. The prokaryotic system mainly uses the improved pET plasmid, and the eukaryotic system mainly uses the pIC9K plasmid series. The verified recombinant plasmid is transferred into the host bacteria. After expression, The target bacteria were obtained by fermentation, and then the bacteria were broken by mechanical method or osmotic pressure method, and the supernatant was collected by centrifugation, and the fusion protein 1 was obtained by purification by hydrophobic chromatography and ion exchange chromatography. The target protein was determined b...

example 2

[0011] According to the protein sequence of fusion protein 3 and the principle of codon preference of hosts in different expression systems, the prokaryotic expression gene sequence and eukaryotic expression gene sequence of fusion protein 1 were designed and synthesized by the method of whole gene synthesis combined with PCR. gene sequence. After sequencing and verification, the target gene is connected into the expression vector. The prokaryotic system mainly uses the improved pET plasmid, and the eukaryotic system mainly uses the pIC9K plasmid series. The verified recombinant plasmid is transferred into the host bacteria, and the target gene is obtained through expression and fermentation. The bacteria are broken by mechanical method or osmotic pressure method, centrifuged to obtain supernatant, and purified by hydrophobic chromatography and ion exchange chromatography to obtain fusion protein 3. The target protein was determined by electrophoresis, HPLC, mass spectrometry,...

example 3

[0013] On the basis of the gene sequence of fusion protein 1, the 4 amino acids Lys-Asp-Glu-Leu at its C-terminus were removed by PCR mutation method and replaced with Arg-Glu-Asp-Leu-Lys, similar to Example 1 The method obtained the fusion protein GnRH-PE40, which is the reference substance. The target protein was determined by electrophoresis, HPLC, mass spectrometry, N-terminal amino acid sequence determination and other methods. Finally, the cell viability test was performed.

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Abstract

The invention provides a series of fusion proteins comprising human luteinizing hormone releasing factor (GnRH) or its mutant and pseudomonas exotoxin (PE) or its mutant. The medicinal purpose of the GnRH is for the highly effective eradication of tumor cells.

Description

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Claims

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Application Information

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Owner BEIJING NORTHLAND BIOTECH
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