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Method for evaluating influence of antigen silencing effect on antibody efficacy and application thereof

An antibody and silencing technology, applied in the field of evaluating the impact of antigen silencing effect on antibody efficacy, can solve problems such as affecting the effect of anti-tumor treatment, achieve broad application prospects and improve the effect of experimental signal window

Pending Publication Date: 2022-04-19
上海药明生物医药有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, certain normal tissue cells, especially red blood cells and lymphocytes, have a high expression of CD47, which will have a more obvious antigen silencing effect on antibody drugs, thereby affecting the therapeutic effect of anti-tumor

Method used

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  • Method for evaluating influence of antigen silencing effect on antibody efficacy and application thereof
  • Method for evaluating influence of antigen silencing effect on antibody efficacy and application thereof
  • Method for evaluating influence of antigen silencing effect on antibody efficacy and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] 1. Antibody acquisition

[0024] The CD47 x TAA bispecific antibody targeting CD47 and tumor associated antigen (Tumor Associated Antigen, TAA) was constructed, expressed and purified by the Protein Science Department of the Biological New Drug R&D Service Business Department of Shanghai WuXi Biotechnology Co., Ltd., in which CD47 and The variable region sequences of TAA were all discovered by the Hybridoma Division of the Biological New Drug R&D Service Business Department of Shanghai WuXi Biotechnology Co., Ltd. Monoclonal antibody targeting CD47 The CD47 monoclonal antibody was constructed, expressed and purified by the Protein Science Department of the Biological New Drug R&D Service Business Department of Shanghai WuXi Biotechnology Co., Ltd. The full-length sequence of the antibody is based on the clone number disclosed in the patent (US9017675) Synthesized for the sequence of 5F9.

[0025] 2. The effect of antigen silencing on the binding of antibodies to cells ...

Embodiment 2

[0034] 1. The effect of antigen silencing on the binding of antibodies to tumor cells

[0035] Such as figure 1 As shown in Table 1, the binding affinity (EC 50 ) is 4.80nM, the binding affinity of CD47 monoclonal antibody to tumor cells (EC 50 ) is 0.36nM. However, under the condition of pre-incubation with whole blood solution, the binding affinity of CD47xTAA double antibody to tumor cells (EC 50 ) was 4.73nM, basically unchanged, indicating that it was not affected by the antigen silencing effect, but the binding affinity of CD47 monoclonal antibody to tumor cells (EC 50 ) was 6.41nM, with a change of about 18 times, indicating that it was affected by a more serious antigen silencing effect. Table 1 shows the effect of the antigen silencing effect of red blood cells on the binding of antibodies to tumor cells.

[0036]

[0037] Table 1

[0038] Additionally, if figure 2 As shown in Table 2, the binding affinity (EC 50 ) is 2.65nM, the binding affinity of CD47 m...

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Abstract

The invention discloses a method for evaluating the influence of an antigen silencing effect on the efficacy of an antibody. The method comprises the following steps: 1, carrying out antigen silencing treatment on the antibody to be detected; and 2, carrying out CFSE dyeing on tumor cells expressing TAA target spots, then carrying out solution incubation on the tumor cells and the antibody subjected to antigen silencing treatment, and analyzing the average fluorescence intensity of the antibody combined on the surfaces of the cells. According to the method, an experimental signal window is improved, the influence of the antigen silencing effect on the antibody effect is remarkably evaluated, meanwhile, the antigen silencing effect aiming at solid tumor cells is more truly simulated, and the method has a wider application prospect in research and development of solid tumor therapeutic antibodies.

Description

technical field [0001] The invention relates to a method for evaluating the efficacy of an antibody, in particular to a method for evaluating the effect of antigen silencing on the efficacy of an antibody. Background technique [0002] Developing therapeutic antibodies against certain protein antigens highly expressed on the surface of tumor cells is a common form of drug development. However, a considerable part of protein antigens that can be made into drugs are expressed not only in tumor cells, but also in blood cells or normal tissue cells. Therefore, when an antibody drug enters the body, it will bind to non-tumor cells such as blood cells or normal tissue cells that express the corresponding antigen, thereby affecting the distribution and elimination of the antibody drug in the body, and limiting the targeting of the antibody drug to the target tissue of the expected effect. This phenomenon is called Antigen Sink. Therefore, it is of great significance to effectivel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/577
CPCG01N33/533G01N33/577
Inventor 邵家骧李婕聂思惟顾继杰
Owner 上海药明生物医药有限公司
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