Vascular Endothelial Growth Factor (VEGF) acceptor 2/CD3 bispecific single-chain antibody

A growth factor receptor, single-chain antibody technology, applied in the direction of anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, antibody, anti-animal/human immunoglobulin, etc., to achieve small molecular weight, immunogen The effect of low performance and high yield

Active Publication Date: 2011-10-19
HARBIN MEDICAL UNIVERSITY
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Problems solved by technology

Bispecific single-chain antibody against VEGF...

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  • Vascular Endothelial Growth Factor (VEGF) acceptor 2/CD3 bispecific single-chain antibody
  • Vascular Endothelial Growth Factor (VEGF) acceptor 2/CD3 bispecific single-chain antibody
  • Vascular Endothelial Growth Factor (VEGF) acceptor 2/CD3 bispecific single-chain antibody

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Embodiment Construction

[0028] 1. Design of bscVEGFR2×CD3 gene fragment and construction of expression vector

[0029] According to VL VEGFR2 , VH VEGFR2 , VH CD3 , VL CD3 The sequence of the amino acid sequence and functional domains VL VEGFR2 -(Gly 4 Ser) 3 -VH VEGFR2 -Gly 4 Ser-VH CD3 -(Gly 4 Ser) 3 -VL CD3. Add the HindIII restriction site, KOZAK sequence and secretion signal peptide sequence in sequence at the 5' end of the sequence, design the KOZAK sequence GCCACCATG before the start codon ATG, which can improve the translation initiation efficiency, and add the secretion signal peptide sequence after ATG (SEQ ID NO: 5) for secretory expression in mammalian cells. Six histidine tags and an EcoR I restriction site were sequentially added to the 3' end of the sequence, and the histidine tags facilitated the purification and identification of recombinant bispecific antibodies. The schematic diagram of the whole gene sequence of bscVEGFR2×CD3 is: KOZAK-Secretary signal sequence-VL V...

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Abstract

The invention relates to a genetic engineering antibody, in particular to a vascular Endothelial Growth Factor (VEGF) acceptor 2/CD3 bispecific single-chain antibody. The bispecific single-chain antibody comprises two binding sites respectively having antigenic specificity on the VEGF acceptor 2 and CD3. The invention further relates to nucleotide sequence coding the bispecific antibody, a carrier containing nucleotide sequence, and a host cell transfected by the carrier. The VEGF2/CD3 bispecific single-chain antibody has small molecular weight and immunogenicity, is easy to penetrate throughcompact tumor barrier and enters the microcirculation around solid tumor, thus having application prospects in preparing tumor treatment drugs.

Description

technical field [0001] The invention relates to a genetic engineering bispecific antibody, in particular to an anti-vascular endothelial cell growth factor receptor 2 / anti-CD3 bispecific single-chain antibody. The present invention also relates to a nucleotide sequence encoding the bispecific antibody, a vector containing the nucleotide sequence, and a host cell transfected with the vector. Background technique [0002] Bispecific single-chain antibody (bsc-Ab) has two antigen-binding arms, which can simultaneously bind to trigger factors on the surface of effector cells and target cells, thereby activating quiescent effector cells and recruiting them around target cells. Mediates lysis of target cells. Usually the mediating effector cells are mainly T cells, whose main function is to identify and rapidly destroy diseased cells or tissues. Activation of T cells is the fundamental premise for cell killing. CD3 molecule is a membrane antigen widely distributed on the surfac...

Claims

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Application Information

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IPC IPC(8): C07K16/28C12N15/11C12N15/85C12N5/10A61K39/395A61P35/00
Inventor 唐晓波李淑珍李郁梅鄢成伟陈莉李新禹
Owner HARBIN MEDICAL UNIVERSITY
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