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Humanized antibody and application thereof

A technology of humanized antibodies and anti-tumor drugs, applied in the direction of antibodies, applications, anti-tumor drugs, etc., to achieve the effect of inhibiting the growth of tumor cells

Active Publication Date: 2022-02-11
PEKING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, apart from monoclonal antibody RP215, there are no other diagnostic or therapeutic drugs that specifically target this IgG.

Method used

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  • Humanized antibody and application thereof
  • Humanized antibody and application thereof
  • Humanized antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Preparation and Screening of Humanized Antibody

[0033] 1. Obtained anti-SIG monoclonal antibody

[0034] Construct a recombinant IgG plasmid with an amino acid sequence as shown in SEQ ID NO.5 of the coding gene of the IgG epitope peptide, and transfer it into the host cell for cultivation. The expression product is verified to have a correct amino acid sequence. N-glycosyl sialic acid modification, which is identified as SIG, is collected and preserved.

[0035] SIG was used as antigen to immunize BALB / c mice. After multiple immunizations, mouse spleen cells were collected, fused with myeloma cells, and hybridoma cells were cultured. Positive hybridoma cells capable of producing anti-SIG antibodies were screened out and cloned and expanded. Then, the anti-SIG monoclonal antibody secreted by it will be identified and sequenced for immunoglobulin type, subclass, specificity, affinity, epitope and molecular weight of the recognized antigen, and hybridoma cell...

Embodiment 2

[0060] Example 2 Functional verification of humanized antibodies

[0061] 2.1 Inhibit cancer cell proliferation and invasion

[0062] Taking lung squamous cell carcinoma cells-NCI-H520 as the experimental object, it was divided into three groups, and 10,000 cancer cells were added to a single well in each group, and each group was replicated three times. The first group was added with mlgG as a negative control, the second group was added with SIG-001, and the third group was added with RP215 as a positive control. Changes in the number of cancer cells in each group. In addition, the invasion ability of the three groups of antibodies was detected by Tran-swell invasion assay.

[0063] The result is as Figure 5 As shown in the above figure, it can be seen that the number of cancer cells in the wells of the culture plate added with antibody SIG-001 is equivalent to that added with RP215, which is significantly less than that of the negative control group added with mlgG, ind...

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PUM

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Abstract

The invention discloses a humanized antibody and application thereof, the humanized antibody comprises a heavy chain variable region and a light chain variable region, and can specifically target a CH1 structural domain 162 site sialylated epitope of IgG, the amino acid sequence of the heavy chain variable region is as shown in SEQ ID NO. 1, and the amino acid sequence of the light chain variable region is as shown in SEQ ID NO. 2. The humanized antibody can effectively inhibit the growth, invasion and suspension growth ability of tumor cells, has an obvious anti-tumor effect, and also can inhibit the activation of a c-Met / Wnt signal channel and an FAK signal channel of the tumor cells.

Description

technical field [0001] The invention relates to the technical field of immunity and genetic engineering, in particular to a humanized antibody and its application in antitumor drugs. Background technique [0002] Previous studies have found that not only B cells highly express IgG, but also non-B cells from a variety of different lineages can also express IgG. Moreover, non-B cell-derived IgG is structurally and functionally different from traditionally understood B cell-derived IgG. There is a big difference. In particular, it was found that the expression level of epithelial tumor cells was significantly higher than that of normal cells. This kind of IgG highly expressed in tumor cells can promote the adhesion ability of tumor cells, promote the migration and invasion of tumor cells in vitro and in vivo, and tumor stem cells are highly The expressed IgG can enhance the self-renewal of tumor stem cells and the ability of tumor formation and metastasis. Based on this, a re...

Claims

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Application Information

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IPC IPC(8): C07K16/42C07K16/44C12N15/13A61K39/395A61P35/00
CPCC07K16/4283C07K16/44A61P35/00C07K2317/56C07K2317/24C07K2317/73A61K2039/505A61K2039/86A61K2039/852A61K2039/82
Inventor 邱晓彦何峙峤黄歆梅姜文华
Owner PEKING UNIV
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