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CD 3 and PRLR bispecific antibody and construction and application thereof

A bispecific antibody and antibody technology, applied in the direction of antibodies, specific peptides, anti-tumor drugs, etc., to achieve obvious effects

Active Publication Date: 2019-04-16
SHANGHAI JIAO TONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above studies imply that PRLR immunotherapy may have minimal serious side effects

Method used

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  • CD 3 and PRLR bispecific antibody and construction and application thereof
  • CD 3 and PRLR bispecific antibody and construction and application thereof
  • CD 3 and PRLR bispecific antibody and construction and application thereof

Examples

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Embodiment 1

[0053] 1. Construction of expression vectors for fragment A and fragment B

[0054] The antibody proteins of fragment A and fragment B bind to CD3 and PRLR extracellular regions respectively, and the amino acid sequences of CD3 and PRLR extracellular regions are shown in Table 1. The expression vectors are respectively according to the antibody A protein structure ( figure 1 ), Fragment A protein consists of two heavy chains (CD3Hc and Int C Fc and a light chain (CD3Lc). Vectors corresponding to the three chains were designed separately. CD3 antibody variable region gene heavy chain and light chain sequence, the required gene segment is synthesized by gene chemistry, and the corresponding gene segments of CD3Hc and CD3Lc protein chains are assembled by PCR amplified oligonucleotide annealing connection, through the restriction site HindIII / BamH1 and other cloning expression vectors, expression vectors can choose mammalian cell expression vectors. In order to solve the hea...

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Abstract

The invention discloses a CD 3 and PRLR duplex-specific antibody and construction and application thereof. A construction method of the CD 3 and PRLR duplex-specific antibody comprises the steps thatheavy and light chain plasmids pM-CD3Hc, pM-CD3Lc and pM-IntCRc of a target CD 3 antibody fusion protein IntC are constructed, subjected to transient cotransfection to be transferred into a cell and subjected to affinity purification, and thus a fragment A antibody is obtained; heavy and light chain plasmids pM-PRLRIntN and pM-PRLRLc of a target PRLR antibody fusion protein IntN are constructed, subjected to transient cotransfection to be transferred into the cell and subjected to affinity purification, and thus a fragment B antibody is obtained; the fragment A antibody and the fragment B antibody are subjected to external trans-splicing, and thus the CD 3 and PRLR duplex-specific antibody is obtained. The CD 3 and PRLR duplex-specific antibody provides a new target therapy strategy for current breast cancer HER 2 and ER target treatment; compared with a PRLR blocking antibody, the PRLR-DbsAb has a more obvious inhibiting effect on tumours.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a CD3 and PRLR bispecific antibody and its construction and application. Background technique [0002] Binding of prolactin to PRLR leads to dimerization of the receptor and intracellular signaling. Signaling through PRLRs has been linked to various processes such as mammary gland development, lactation, reproduction and immune regulation. Locally produced prolactin, in addition to prolactin produced by the anterior pituitary, can promote tumorigenesis in the autocrine form of breast cancer. PRLR is slightly expressed in breast tissue in normal tissue, and the expression level in breast cancer tissue is 30 times higher than that in normal breast tissue. In a study of two orthotopic and highly metastatic breast cancer cell lines (mouse 4T1 and human BT-474), knockdown of PRLR in 3D culture resulted in 95% tumor initiation / apoptosis of cancer stem cells ( Yonezawa, T., e...

Claims

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Application Information

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IPC IPC(8): C07K16/46C12N15/85A61K39/395A61P35/00
CPCA61K2039/505A61P35/00C07K16/2809C07K16/2869C07K16/468C07K2317/31C12N15/85
Inventor 朱建伟周跃鲜宗会芳谢跃庆江华韩雷
Owner SHANGHAI JIAO TONG UNIV
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