Anthropogenic LMP1 (latent membrane protein1) extracellular regional antibody for nasopharyngeal carcinoma and application thereof

A technology for the extracellular region and nasopharyngeal carcinoma, which is applied in the field of genetic engineering and immune-targeted diagnosis and treatment, and can solve the problem of lack of human anti-nasopharyngeal carcinoma LMP1 extracellular region antibody

Inactive Publication Date: 2013-05-29
THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is no human-derived antibody against the extracellular domain of LMP1 in nasopharyngeal carcinoma

Method used

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  • Anthropogenic LMP1 (latent membrane protein1) extracellular regional antibody for nasopharyngeal carcinoma and application thereof
  • Anthropogenic LMP1 (latent membrane protein1) extracellular regional antibody for nasopharyngeal carcinoma and application thereof
  • Anthropogenic LMP1 (latent membrane protein1) extracellular regional antibody for nasopharyngeal carcinoma and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation and Identification of Human Anti-LMP1 Extracellular Region Antibody Fab

[0031] 1. Preparation and purification of latent membrane protein 1 (LMP1) extracellular region antigen

[0032] 1. The gene sequence of the extracellular region of LMP-1 comes from the data published by GenBank. The three extracellular regions of LMP-1 LMP-1[46-50]DWTGG, [98-103]WNLHGQ, and [161-166]QQNWW are spliced ​​into WNLHGQ WNLHGQ QQNWW QQNWW DWTGG WNLHGQ WNLHGQ QQNWW QQNWW DWTGG (SEQ ID NO.5); gene sequence is 5'-GC GGATCC TGG AAC CTG CAC GGT CAG TGG AAC CTG CAT GGT CAA CAG CAA AAC TGG TGG CAA CAA AAC TGG TGG GAC TGG ACC GGT GGC TGG AAC CTG CAC GGT CAG TGG AAC CTG CAC GGC CAG CAG CAA AAC TGG TGG CAG CAG AAC TGG TGG GAT TGG ACT GGC GGT TGA GAATTC G-3' (SEQ ID NO.6); and the restriction sites (underlined) of BamH I and EcoR I were designed at the 5' end and 3' end respectively, and the whole gene was synthesized by Nanjing Saibaisheng Biological Co., Ltd., and Th...

Embodiment 2

[0210] Example 2: Preparation of HLEAFab-MMC immunotoxin and its in vivo and in vitro effects on nasopharyngeal carcinoma cells

[0211] 1. Preparation of HLEAFab-MMC immunotoxin

[0212] 1. MMC was reacted with n-hydroxysuccinimide 3-(2-pyridinedimercapto)propionate (SPDP, Pierce, Rockford, IL) at 4°C for 2 hours, and then placed at -80°C for use.

[0213] 2. HLEAFab was reduced with 2-iminosulfane hydrochloride in pure nitrogen, and the reduced antibody was purified using a desalting column (Pierce, Rockford, IL).

[0214] 3. The activated HLEAFab and the activated mitomycin (Mytomycin C, MMC) were mixed and reacted (HLEAFab:Mytomycin C molar ratio=1:1). React for 1 hour at room temperature in pure nitrogen, and remove excess MMC-SPDP using a desalting column.

[0215] 4. HLEAFab-MMC products are sterilized by filtration with a 0.2 μm ultrafiltration membrane (Pall, Ann Arbor, MI).

[0216] 5. Cell ELISA (the specific steps are the same as the HLEAFab cell ELISA detection...

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Abstract

The invention belongs to the fields of genetic engineering and immunity-targeted diagnosis and treatment and discloses an anthropogenic LMP1 (latent membrane protein1) extracellular regional antibody for nasopharyngeal carcinoma and application thereof. The amino acid sequence of the variable region of the light chain of the antibody is shown as SEQ ID NO.1, the amino acid sequence of the variable region of the heavy chain of the antibody is shown as SEQ ID NO.2, and the antibody can be combined with a human nasopharyngeal carcinoma LMP1 extracellular region and can be used in the preparationof drugs acting on the human nasopharyngeal carcinoma LMP1 extracellular region in a targeting manner.

Description

Technical field: [0001] The invention belongs to the field of genetic engineering and the field of immune targeting diagnosis and treatment, and relates to a human anti-nasopharyngeal carcinoma LMP1 extracellular region antibody and application thereof. Background technique: [0002] Nasopharyngeal carcinoma is a highly malignant tumor derived from the nasopharyngeal epithelium. The tumor progresses and easily invades important structures such as the skull base, and early cervical lymph node metastasis and distant metastasis occur. Nasopharyngeal carcinoma is characterized by ethnic and regional distribution. It is one of the top ten high-incidence malignant tumors in my country. Its incidence rate is 20 / 100,000, which has caused serious health problems. Nasopharyngeal carcinoma (NPC) is a complex disease gradually formed by the multi-step interaction involving latent infection of Epstein-Barr virus (EBV), environmental factors and genetic factors of the host. All nasophary...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/08C12N15/13C12N15/63A61K47/48A61K31/407A61P35/00
Inventor 陈仁杰张大为朱进冯振卿
Owner THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV
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