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VEGF-D (vascular endothelial growth factor-D) monoclonal antibody and kit

A VEGF-D, monoclonal antibody technology, applied in the biological field, to achieve the effect of simple detection method, low cost, and good biological activity

Inactive Publication Date: 2018-11-13
浙江众意生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In the prior art, the biomarker products for tumor lymphatic metastasis are still blank in China, and there is still the possibility of further improvement in the specificity and affinity of similar products. Based on this, we have developed a VEGF-C assay kit (chemiluminescence method) and its preparation method

Method used

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  • VEGF-D (vascular endothelial growth factor-D) monoclonal antibody and kit
  • VEGF-D (vascular endothelial growth factor-D) monoclonal antibody and kit
  • VEGF-D (vascular endothelial growth factor-D) monoclonal antibody and kit

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Experimental program
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Embodiment 1

[0022] 1. Preparation of VEGF-D antigen:

[0023] (1) Design a pair of primers according to the sequence of human VEGF-D (D89630.1) in the GenBank database. The upstream primer is vP: 5'-ATATGTACAGAGAGTGGGTAGTGGTGAATG-3', and the XhoI restriction site is introduced; the downstream primer is vP: 5' -TTAACAGCATGCTGCTTTGC-3', introduce EcoRI restriction site, extract total RNA from human hepatocytes by RT-PCR method, then reverse transcribe into cDNA, use cDNA as template, use Pyrobest DNA polymerase for VEGF-D gene specificity sexual amplification. The amplified PCR product was recovered from the gel, and the recovered VEGF-D gene and pPIC9K plasmid were respectively digested with Xho I and EcoR I, and the recovered target fragment was ligated with T4 ligase, and the ligated product was transformed into DH5α competent cells, passed through ampicillin (1 mg / mL) antibody screened positive clones, and the plasmids were extracted and identified by enzyme digestion and sequencing. ...

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Abstract

The invention discloses a VEGF-D (vascular endothelial growth factor-D) monoclonal antibody. The amino acid sequence of a heavy chain of the VEGF-D monoclonal antibody is as shown in VH:BAA24264, andthe amino acid sequence of a light chain of the VEGF-D monoclonal antibody is as shown in VL:XP_009304362. The VEGF-D monoclonal antibody has high affinity, strong specificity and good biological activity in vitro. A VEGF-D protein detection kit built by the VEGF-D monoclonal antibody and a VEGF-D polyclonal antibody can serve as an index for evaluating tumor lymphangiogenesis and lymphatic metastasis.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a VEGF-D monoclonal antibody and a VEGF-D protein detection kit. Background technique [0002] In the study of the mechanism of tumor metastasis, the importance of the lymphatic system as a channel for the metastasis of tumors, especially solid tumors, has been continuously recognized. Clinicopathological studies have shown that the earliest route of dissemination of solid tumors is regional lymph node dissemination via lymphatic channels. At present, most studies believe that the VEGF-D / VEGFR-3 signaling system is the most important regulatory mechanism of tumor lymphangiogenesis, and plays an important role in the process of tumor lymphatic metastasis. [0003] VEGF-D is the 1998 Achen et al. [1] It was found by computer homology search, similar to VEGF-C structure, located on chromosome Xp22.31. VEGF-D binds to endothelial cell receptors in humans, and proteolytic p...

Claims

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Application Information

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IPC IPC(8): C07K16/22C12N15/13C12N15/63G01N33/68G01N33/577
CPCC07K16/22C07K2317/56C12N15/63G01N33/577G01N33/6872G01N2333/475
Inventor 吕鹏辉
Owner 浙江众意生物科技有限公司
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