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A novel process for purification of human growth hormone

A technology of human growth hormone, hydrophobic interaction chromatography, applied in a new field for purification of human growth hormone

Inactive Publication Date: 2007-10-24
USV LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0021] A limitation of the current technology is the use of a non-ionic surfactant, Brij 35, to improve process recovery at analytical levels

Method used

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  • A novel process for purification of human growth hormone
  • A novel process for purification of human growth hormone
  • A novel process for purification of human growth hormone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Addition of K to the ion-exchange purified fraction of hGH 2 HPO 4 The final concentration is 0.4M. Inject it onto a 1ml Resource phenyl column (30×6.4mm) with 20mM Na 2 HPO 4 / 0.4M K 2 HPO 4 , pH9.0 balance. Bind protein with 20ml linear gradient of 20mM Na 2 HPO 4 , pH 9.0 / 50% acetonitrile for elution. Fractions of 1 ml were collected and analyzed by SDS-PAGE. The clipped hGH molecules elute before intact GH molecules (Figure 1). Obtain human growth hormone in a highly pure form. (purity: >99.5%, yield: 85%)

Embodiment 2

[0087] Addition of K to the ion-exchange purified fraction of hGH 2 HPO 4 The final concentration is 0.4M. Inject it onto a 1ml Resource phenyl column (30×6.4mm) with 20mM Na 2 HPO 4 / 0.4M K 2 HPO 4 , pH9.0 balance. Bind protein with 20ml linear gradient of 20mM Na 2 HPO 4 , pH 9.0 / 40% acetonitrile for elution. Fractions of 1 ml were collected and analyzed by SDS-PAGE. The clipped hGH molecules elute before intact GH molecules (Figure 2). Obtain human growth hormone in a highly pure form. (purity: >99.5%, yield: 85%)

Embodiment 3

[0089] Addition of K to the ion-exchange purified fraction of hGH 2 HPO 4 The final concentration is 0.4M. Inject it into a 1ml Resource phenyl column (30×6.4mm) with 20mM Na 2 HPO 4 / 0.4M K 2 HPO 4 , pH9.0 balance. Bind protein with 20ml linear gradient of 20mM Na 2 HPO 4 , pH 9.0 / 30% acetonitrile for elution. Fractions of 1 ml were collected and analyzed by SDS-PAGE. This clipped hGH molecule was not resolved from the intact molecule (Figure 3).

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Abstract

A substrate for growth of nitride semiconductor capable of obtaining a high-quality nitride semiconductor crystal layer is provided. A substrate for growth of nitride semiconductor for growth of a nitride semiconductor layer on a sapphire substrate ( 1 ) according to one embodiment of the invention is provided with an Al2O3 layer ( 2 ) as separately provided on the sapphire substrate ( 1 ), an AlON layer ( 3 ) which is the first layer, and an AlN layer ( 4 ) which is the second layer. With respect to the first layer and the second layer, the AlON layer ( 3 ) and the AlN layer ( 4 ) are deposited on the Al2O3 layer ( 2 ) in this order.

Description

technical field [0001] The present invention relates to a method of purifying human growth hormone (hGH) on a commercial scale by using hydrophobic interaction chromatography, which efficiently separates clipped hormone fractions formed during the production of growth hormone by recombinant DNA techniques. Background technique [0002] Human growth hormone (hGH) is a pituitary-derived protein that has several important biological functions, including protein synthesis, cell proliferation, and metabolism. hGH is a 191 amino acid residue polypeptide of approximately 22 kDa. Recombinant human growth hormone (hGH) has been expressed in E. coli as an intracellular and secreted protein. A series of chromatographic and / or non-chromatographic methods have been used to obtain pure proteins. The region between amino acid residues 140-150 in human growth hormone has been reported to be sensitive to various proteases. This results in proteolytically cleaved forms of the protein whose...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/61C07K1/20
CPCC07K1/20C07K14/61
Inventor 阿麦德·蒙索尔·博尔伯惠亚亚达·麻德哈瓦·劳拉克斯米·斯利尼瓦斯·劳帝潘威塔·麦提普拉布哈卡尔·米林德·尼普哈德卡尔施利康德·米施拉
Owner USV LTD