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Biological activity determination method for anti-PD-L1 monoclonal antibody

A PD-L1 and monoclonal antibody technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of rising patient mortality and enhancing tumor metastasis, and achieve good durability, strong specificity, precision and The effect of high accuracy

Active Publication Date: 2016-06-29
兆科(广州)肿瘤药物有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In tumor patients, high expression of PD-L1 can enhance tumor metastasis and lead to increased mortality

Method used

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  • Biological activity determination method for anti-PD-L1 monoclonal antibody
  • Biological activity determination method for anti-PD-L1 monoclonal antibody
  • Biological activity determination method for anti-PD-L1 monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 Determination of the biological activity of 6 different batches of anti-PD-L1 monoclonal antibodies

[0074] The 6 batches of different batches of anti-PD-L1 monoclonal antibodies are anti-PD-L1 monoclonal antibodies produced by our company, and the determination steps are as follows:

[0075] (1) Take out the frozen PD1 cells and PD-L1 cells from the liquid nitrogen tank, thaw and recover the cells quickly, transfer them to a culture bottle containing cell growth medium, and store them at 37°C, 5% CO 2 cultured in an incubator.

[0076] (2) Take PD-L1 cells in the logarithmic growth phase, digest, centrifuge, and dilute the cells to 4×10 5 cells / mL, and then inoculated in a 96-well plate at a seeding density of 4×10 per well 4 at 37°C, 5% CO 2 Incubate overnight in an incubator.

[0077] (3) Prepare 6 batches of different batches of anti-PD-L1 monoclonal antibody sample solutions, measure their concentration by BCA method, and dilute the samples to be tes...

Embodiment 2

[0084] Example 2 Using anti-PD-L1 monoclonal antibody, methodological verification of this method

[0085] Except for the special instructions in the following items, the experimental materials and methods were basically the same as those in Example 1.

[0086] 1 specificity

[0087] In this experiment, the specificity of target cells, effector cells and anti-PD-L1 monoclonal antibodies were investigated respectively. Specifically, they are:

[0088] 1.1 Target cell specificity: the effector cells are Jurkat / PD1, the antibody is anti-PD-L1 monoclonal antibody, and the target cells are CHO / PD-L1, CHO / PD-L1 negative cell and Dilutionbuffer.

[0089] 1.2 Effector cell specificity: the target cell is CHO / PD-L1, the antibody is anti-PD-L1 monoclonal antibody, and the effector cells are Jurkat / PD1, Jurkat / PD1negativecell and Dilutionbuffer.

[0090] 1.3 Specificity of anti-PD-L1 monoclonal antibody: the target cell is CHO / PD-L1, the effector cell is Jurkat / PD1, and the antibodies...

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Abstract

The invention relates to the field of activity detection of biological medicine, in particular to a biological activity determination method for an anti-PD-L1 monoclonal antibody. According to the method, a PD1 / PD-L1 interruption detection system is used, binding of PD1 and PD-L1 protein is interrupted when the anti-PD-L1 monoclonal antibody is bound with PD-L1 expressed by target cells, a transcription factor NFAT is activated through signal transduction, expression of luciferase is started, the expression quantity of luciferase is positively correlated to the biological activity of the anti-PD-L1 monoclonal antibody, the chemiluminescence value of the anti-PD-L1 monoclonal antibody is determined after a cell lysis solution and a luciferase substrate are added, a dose-effect curve is drawn, and accordingly the biological activity of the anti-PD-L1 monoclonal antibody is determined. The method is easy to implement, strong in specificity and high in durability and has great significance on quality control and clinical application of the anti-PD-L1 monoclonal antibody.

Description

technical field [0001] The invention relates to the field of biological drug activity detection, in particular to a method for measuring the biological activity of an anti-PD-L1 monoclonal antibody. Background technique [0002] The PD-1 / PD-L1 signaling pathway is closely related to the immune escape of tumors and viruses, and is an important new target for immunotherapy of tumors and chronic infectious diseases. Using monoclonal antibodies against PD-1 and PD-L1 to block the PD-1 / PD-L1 signaling pathway and restore the immune killing function of T cells can exert a good therapeutic effect and has a good application prospect. [0003] PD-1 is an immune co-inhibitory molecule, which belongs to the CD28 family member. It is a type I transmembrane protein composed of 268 amino acids and located on the PDCD1 gene. Its structure mainly includes the extracellular immunoglobulin variable region (IgV) domain, a hydrophobic transmembrane region, and an intracellular region. The int...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 梁杰姚菲黄思伽戴向荣李小羿
Owner 兆科(广州)肿瘤药物有限公司
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