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Method for detecting biological activity of recombinant human interleukin-1 receptor antagonist eye drops

A technology of interleukin and receptor antagonist, applied in the field of detection of biological activity of recombinant human interleukin 1 receptor antagonist eye drops, can solve the problems of many detection steps, poor specificity, unstable results, etc. Achieve the effect of good repeatability, simple operation and easy observation

Active Publication Date: 2017-08-22
西藏诺迪康药业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] But there are many deficiencies in the above method, such as PGE 2 There are many detection steps; the mouse thymocyte method is very unstable due to individual differences in the living body; other T lymphocytes not only have a proliferation effect on IL-1, but also have an effect on other cytokines, and the specificity is poor

Method used

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  • Method for detecting biological activity of recombinant human interleukin-1 receptor antagonist eye drops
  • Method for detecting biological activity of recombinant human interleukin-1 receptor antagonist eye drops
  • Method for detecting biological activity of recombinant human interleukin-1 receptor antagonist eye drops

Examples

Experimental program
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Effect test

Embodiment 1

[0051] Preparation of standard solution: Dissolve 10 μg of rhIL-1ra standard (1U / mg, NIBSC) in 1.25ml of water, aliquot and freeze, and use culture medium B before use 2 Dilute 10 times to make the concentration 800ng / ml. In a 96-well culture plate, make a 3-fold serial dilution, a total of 11 dilutions, make 3 wells for each dilution, leave 50μl standard solution in each well, discard excess solution in the well. The above operations were carried out under sterile conditions.

[0052] Preparation of the test solution: get the rhIL-1ra eye drops prepared above, add culture solution B 2 , until the concentration of rhIL-1ra is 800ng / ml, in a 96-well culture plate, make 3-fold serial dilutions, a total of 11 dilutions, each dilution is made into 3 holes, and 50 μl of the test solution is left in each hole, discarded excess solution in the well. The above operations were carried out under sterile conditions.

[0053] Assay:

[0054] Add the A375-s2 cell suspension to each we...

Embodiment 2

[0065] Preparation of standard solution: Dissolve 10 μg of rhIL-1ra standard (1U / mg, NIBSC) in 1.25ml of water, aliquot and freeze, and use culture medium B before use 2 Dilute 10 times to make the concentration 800ng / ml. In a 96-well culture plate, make a 3-fold serial dilution, a total of 11 dilutions, make 2 wells for each dilution, leave 50μl standard solution in each well, discard excess solution in the well. The above operations were carried out under sterile conditions.

[0066] Preparation of the test solution: get the rhIL-1ra eye drops prepared above, add culture solution B 2 , until the concentration of rhIL-1ra is 800ng / ml, in a 96-well culture plate, make 3-fold serial dilutions, a total of 11 dilutions, each dilution is made into 2 wells, and 50 μl of the test solution is left in each well, discarded excess solution in the well. The above operations were carried out under sterile conditions.

[0067] Assay:

[0068] Add the A375-s2 cell suspension to each we...

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Abstract

The invention discloses a method for detecting the biological activity of recombinant human interleukin-1 receptor antagonist eye drops. The method comprises the following steps: (1) preparing a standardized products solution; (2) preparing a solution for test products; (3) measuring absorbance, recording the measurement result, processing test data by a computer program or four parameter regression calculation method, and calculating the biological activity of a test product. The method for detecting the biological activity of recombinant human interleukin-1 receptor antagonist eye drops is simple to operate and easy for observation, has good linear relation, good repeatability and strong specificity, and provides an effective technological means for monitoring product quality.

Description

technical field [0001] The invention belongs to the field of detection of biopharmaceutical activity, and in particular relates to a detection method of biological activity of recombinant human interleukin-1 receptor antagonist eye drops. Background technique [0002] IL-1ra can competitively bind to the receptor of IL-1 on T lymphocytes, thereby inhibiting the biological activity of IL-1. Based on this principle, a variety of detection methods for the biological activity of IL-1ra have been established: IL-1ra 1ra inhibits IL-1-induced proliferation of mouse thymocytes; IL-1ra inhibits IL-1-induced PGE secretion from human dermal fibroblasts 2 Experiment and inhibit other T lymphocyte lines such as D10AG4.1 and D10(N 4 ) proliferation experiments. [0003] But there are many deficiencies in the above method, such as PGE 2 There are many detection steps; the mouse thymocyte method is very unstable due to individual differences in the living body; other T lymphocytes not o...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N21/31
CPCG01N21/31G01N33/5008
Inventor 任东升何晨周菊萍冯海燕刘希望张艳
Owner 西藏诺迪康药业股份有限公司
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