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High-stability PD-L1 antibody reagent and application thereof

A PD-L1, high-stability technology, applied in the field of molecular biology, can solve the problems of poor antibody stability, decreased titer, inconvenient storage and access, etc. Avoid the effect of antibody titer reduction

Active Publication Date: 2019-06-04
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the traditional primary antibody reagents used in immunohistochemistry or other immunological experiments need to be stored at -20°C, and dilution steps are required before use, making storage and access more inconvenient.
At the same time, it needs to be dissolved when stored at -20°C. Repeated freezing and thawing will deteriorate the stability of the antibody and easily lead to the problem of decreased titer.

Method used

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  • High-stability PD-L1 antibody reagent and application thereof
  • High-stability PD-L1 antibody reagent and application thereof
  • High-stability PD-L1 antibody reagent and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: PD-L1 immunohistochemistry kit is used to detect the expression of PD-L1 in NSCLC tissue sections

[0043] This embodiment provides an immunohistochemical kit for detecting PD-L1 and its application. The kit includes the following reagents:

[0044] Endogenous peroxidase blocker, 3% (v / v) H 2 o 2 solution;

[0045] Antibody reagent with high stability: mouse anti-human PD-L1 antibody PD-L1 Monoclonal Antibody, (ThermoFisher: 14-5983-80), diluted to 0.5 μg / ml with Peroxidase Stabilizing Buffer;

[0046] HRP-goat anti-mouse IgG, purchased from Maixin Biotechnology Development Co., Ltd., Kit-0014;

[0047] Blocking solution, 5% (w / v) bovine serum albumin (BSA) solution prepared in phosphate buffer saline PBS as a solvent;

[0048] DAB chromogenic solution was purchased from Maixin Biotechnology Development Co., Ltd.

[0049] The detection principle of the kit is as follows: the immunological antigen and the antibody can be specifically combined based on the...

Embodiment 2

[0081] Embodiment 2: Antibody stabilizer affects research on antibody stability

[0082] Use Sigma’s Peroxidase Stabilizing Buffer (Cat. No. P 9209), CANDOR Bioscience’s Antibody Stabilizer PBS (Cat. No.: 131500) and HRP-Stab (Catalog No.: 270500) was used as an antibody stabilizer to dilute mouse anti-human PD-L1 Monoclonal Antibody, (Thermo Fisher: 14-5983-80), diluted to 0.4 μg / ml, and each antibody stabilizer was tested by accelerated stability test Ability to stabilize monoclonal antibody titers. The diluted primary antibody was stored at 37°C, and immunohistochemical detection was performed on B-CPAP and MCF-7 cells with Maixin DAB detection kit on day 0, day 4, day 7, and day 9, respectively, and the staining effect was compared. Store the mouse anti-human PD-L1 Monoclonal Antibody diluted in PBS, (Thermo Fisher: 14-5983-80) at 37°C for 0, 4, 7, and 9 days, and dilute it with PBS on the same day, and prepare and use the mouse anti-human PD -L1 Monoclonal Antibody, (T...

Embodiment 3

[0088] Example 3: Influence of antibody concentration on the detection effect of immunohistochemistry

[0089] Dilute at different concentrations:

[0090] The PD-L1 antibody stock solution described in Example 1 was 0.5 mg / ml, and was diluted with Peroxidase Stabilizing Buffer (Cat. No. P 9209) to the following concentrations: 0.8 μg / ml, 0.6 μg / ml, 0.4 μg / ml and 0.2 μg / ml. ml. The antibody solutions of the above 4 concentrations and the anti-human PD-L1 antibody (Cat. No. MAB1561) purchased by R&D were used as controls, and were used for immunohistochemical experiments on the 0th, 4th, 7th, and 9th days at room temperature, and the effect was determined.

[0091] In this example, the cultured thyroid cancer cell line B-CPAP and human breast cancer cell line MCF-7 formaldehyde-fixed paraffin-embedded tissue sections were used as samples, and the Maixin DAB kit was used to conduct immunohistochemical tests to test PD-L1 monocytogenes. The titer of the cloned antibody was used...

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Abstract

The invention discloses a high-stability PD-L1 antibody reagent, which comprises a mouse anti-human PD-L1 monoclonal antibody and an antibody stabilizer, wherein the antibody stabilizer is selected from a group consisting of Peroxidase Stabilizing Buffer and Sigma: P9209 (Item No. P 9209); the mouse anti-human PD-L1 monoclonal antibody is selected from a group consisting of PD-L1 Monoclonal Antibody and Thermo Fisher: 14-5983-80. The high-stability PD-L1 antibody reagent provided by the invention obtains the antibody which can be stably stored at 4 DEG C through the reasonable cooperation of the antibody stabilizer and the PD-L1 monoclonal antibody and is convenient to store, avoids the reduction of antibody valence caused by repeated freezing and thawing, and integrates reagents requiredfor the immunohistochemical detection of PD-L1 by means of a provided reagent kit, thereby making the immunohistochemical detection of PD-L1 more intuitive, more convenient and faster.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a PD-L1 immunohistochemical kit. Background technique [0002] Lung cancer is one of the most common malignant tumors in the world, and its morbidity and mortality are at the end of tumors, and there is an increasing trend year by year. Non-small cell lung cancer (NSCLC) accounts for about 80-85% of the total number of lung cancers, and about 75% of the patients were found in the middle and advanced stages. The average survival time was 12.9 months, and the 3-year and 5-year survival rates were higher. low, at 19% and 11%, respectively. At present, the treatment methods of non-small cell lung cancer mainly include surgery, chemotherapy, radiotherapy and molecular targeted therapy. Surgery remains the only treatment that can potentially cure patients, yet many patients with non-small cell lung cancer are often found out of surgery when they are discovered. In recent years, some...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/574G01N33/68
Inventor 董艳许嘉森吴诗扬刘苏燕
Owner SUREXAM BIO TECH
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