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Novel anti-CD20 antibody nanometer array and its preparation method and use

A CD20, antibody technology, applied in the direction of antibody, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-tumor drugs, etc. The effect of antitumor effect

Inactive Publication Date: 2016-07-06
SHANGHAI ZHANGJIANG BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Novel antibody nanoarrays designed by the present invention have no similar research reports at home and abroad

Method used

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  • Novel anti-CD20 antibody nanometer array and its preparation method and use
  • Novel anti-CD20 antibody nanometer array and its preparation method and use
  • Novel anti-CD20 antibody nanometer array and its preparation method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0049] Example: Preparation method of anti-CD20 antibody nanoarray

[0050] (1) Prepare 4mg / ml PEI (polyethyleneimine, molecular weight 7500kDa) and MPEGS (MAL-PEG-SCM, maleimide-polyethylene glycol-succinimide acetate) solutions respectively, and PEI and MPEGS were mixed at a mass ratio of 1:20. After fully reacting at room temperature for 4 hours, they were dialyzed with a 3500MCW dialysis membrane for 4-6 hours to remove unreacted free MPEGS.

[0051] (2) Thiolated antibody: Take 1mg of Rituximab and 11B8 antibody stock solutions and dilute them to 2mg / ml respectively. According to 2-IT / mAbs=0.15:1, add 5 mg / ml 2-IT (2-iminothiolane) to the antibody solution respectively, place it at room temperature for 2-2.5 hours and then dialyze it with PBS solution containing 5mM EDTA for 6-8 hours. Remove unreacted free 2-IT (dialysis membrane 1000MCW).

[0052] (3) Mix the same amount of thiolated Rotuximab with 11B8, add the MPEGS-PEI solution (mAb / PEI=1000:3.44) prepared in (1) t...

experiment example 1

[0055] Experimental example 1: SDS-PAGE identification of anti-CD20 antibody nanoarray

[0056] The prepared anti-CD20 antibody nanoarray was subjected to SDS-PAGE electrophoresis with 8% separating gel, and then stained with Coomassie brilliant blue to identify the molecular weight of the anti-CD20 antibody nanoarray. SDS-PAGE map see image 3 , where lanes 3 and 4 are anti-CD20 antibody nanoarrays (PL-RB), lane 5 is the antibody Rituximab, and lane 6 is the antibody 11B8.

experiment example 2

[0057] Experimental Example 2: Particle Size Measurement and Morphological Observation of Anti-CD20 Antibody Nanoarray

[0058] After the prepared anti-CD20 antibody nanoarray was diluted with sterilized PBS, the particle size was detected by a dynamic tube scattering instrument (ALV / CGS-3, Germany). The results are shown in Figure 4 .

[0059] Put the diluted sample on the special copper grid for transmission electron microscope, after air-drying, carry out negative staining with 2% PTA (phosphotungstic acid) solution, and observe its morphology with transmission electron microscope. The diluted sample can also be placed under an atomic force microscope for morphological observation by conventional methods.

[0060] See Figure 5 and Image 6 , where the scale bars are 1.5 μm and 500 nm, respectively.

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Abstract

The invention relates to a novel anti-CD20 antibody nanometer array (PL-RB) and its preparation method and a research of novel anti-CD20 antibody nanometer array antineoplastic effects. The novel anti-CD20 antibody nanometer array utilizes long-chain polyethyleneimine as a carrier and realizes in-vitro crosslinking of a type I anti-CD20 antibody Rituximab and a type II anti-CD20 antibody 11B8 so that the novel anti-CD20 antibody nanometer array is prepared. In an in-vitro experiment, the novel anti-CD20 antibody nanometer array successfully activates all known four CD20 antibody antineoplastic paths so that antineoplastic effects are improved.

Description

technical field [0001] The invention relates to a preparation method of an anti-CD20 antibody nano-array and its anti-tumor effect. Background technique [0002] Lymphoma is a malignant tumor that originates in the lymphatic system, and it often occurs in young adults. Epidemiological data show that about 90% of them are non-Hodgkin's lymphoma (NHL), of which about 85% are derived from B cells . In recent years, the incidence of NHL has increased year by year. Although NHL is sensitive to traditional radiotherapy and chemotherapy, it is very easy to relapse, and it is resistant to radiotherapy and chemotherapy after recurrence. [0003] Molecular targeted therapy refers to treatments aimed at cell signal transduction and other biological pathways involved in tumorigenesis and development. The targets of molecular targeted therapy in a broad sense include any submolecules from DNA to protein levels involved in multiple processes such as cell differentiation, apoptosis, mig...

Claims

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Application Information

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IPC IPC(8): C07K16/28A61K39/395A61P35/00
Inventor 钱卫珠
Owner SHANGHAI ZHANGJIANG BIOTECH
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