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Monoclonal antibody for resisting human osteogenesis induction factor and its preparation method and uses

A monoclonal antibody, inducing factor technology, applied in the field of immunology

Inactive Publication Date: 2007-04-25
RUIJIN HOSPITAL ATTACHED TO SHANGHAI NO 2 MEDICALUNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no one in the prior art has obtained specific monoclonal antibodies against human OIF

Method used

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  • Monoclonal antibody for resisting human osteogenesis induction factor and its preparation method and uses
  • Monoclonal antibody for resisting human osteogenesis induction factor and its preparation method and uses
  • Monoclonal antibody for resisting human osteogenesis induction factor and its preparation method and uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Preparation of monoclonal antibodies

[0060] In this example, the inventors used the E. coli system to express and purify OIF, and further prepared OIF-specific monoclonal antibody OIF-KG4, which provided convenience for the further establishment of a detection method for OIF in blood, and was an OIF antibody It is possible to diagnose special types of tumors and lay the foundation for further in-depth study of the physiological function of OIF. The specific process is as follows:

[0061] 1. Materials

[0062] (1) Plasmids, strains and main preparations

[0063] Escherichia coli BL21 and Top10 were purchased from Amersham biosciences. Primer Forward 5′-atc was designed according to AF100758 gga tcc cct gtc aac gca act ctg g-3' (SEQ ID NO: 5), Reverse 5'-atg ctc gag cat tgg ttg agt cct ggg ac-3' (SEQ ID NO: 6), the OIF coding sequence was amplified by PCR, digested with BamH I and Xho I, and cloned into the prokaryotic expression vector pGEX-5X-2 (purchased fro...

Embodiment 2

[0092] Detection of OIF

[0093] Detection by immunohistochemical method. Specifically, New Zealand white rabbits were immunized with GST-OIF protein, and their serum was used as the polyantiserum against OIF and used as the primary antibody. The secondary antibody envision reagent was purchased from DAKO Company. Color was developed with DAB (DAKO company).

[0094] The expression of OIF in 178 cases of lung cancer tissues was detected, and it was found that the expression of OIF was related to the degree of differentiation: the higher the differentiation, the higher the expression. In squamous cell carcinoma tissues, the expression of OIF was almost 100% (Table 3), and the positive rate of SCLC with poor differentiation was the lowest (Table 4). It can be seen that OIF, as a tumor marker of lung cancer, can effectively distinguish small cell lung cancer (SCLC) from non-small cell lung cancer (NSCLC). Helpful for clinicopathological classification.

[0095] ...

Embodiment 3

[0100] Detection of OIF expression with monoclonal antibody OIF-KG4

[0101] The present inventors used the prepared monoclonal antibody OIF-KG4 to detect the expression of OIF, and repeated the method of Example 2, with the difference that the polyclonal antibody in Example 2 was replaced by the monoclonal antibody OIF-KG4 as the primary antibody.

[0102] The results showed that monoclonal antibody OIF-KG4 can also effectively differentiate small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). Moreover, the specificity of the monoclonal antibody OIF-KG4 was greatly improved compared with the polyclonal antibody in Example 2, and the affinity was also significantly enhanced.

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Abstract

The invention discloses an astopic monoclonal antibody OIF-KG4 of osteoinductive factor (OIF) protein and OIF-KG4 immune globulin, segment and immune coupler and drug component, which also provides an agent box to test tumour, obesity, diabetes, hypertension and hyperlipemia.

Description

technical field [0001] The invention belongs to the field of immunology and relates to a monoclonal antibody. More specifically, the invention relates to an anti-human osteogenesis-inducing factor monoclonal antibody, a preparation method and a pharmaceutical composition thereof. Background technique [0002] Osteogenesis-inducing factor (osteoinductive factor OIF), also known as Osteoglycin or Mimecan, is a secreted protein that was first isolated from bovine bone matrix as a 12kDa protein (Bentz H et al., J Biol Chem 1989; 264(34 ):20805-10), and its amino acid sequence was obtained by amino acid sequencing (Bentz H et al., J Biol Chem 1990; 265(9):5024-9). Subsequently, people cloned the human OIF gene from the cDNA library of osteoblasts (Madisen L et al., DNA Cell Biol 1990; 9(5): 303-9), but the cDNA sequence was not published. The human OIF gene encodes a 298-amino acid secretory protein propeptide, which is 86% homologous to bovine OIF. The 1-19th position is the si...

Claims

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Application Information

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IPC IPC(8): C07K16/18C07K16/24C07K14/51C12N5/18G01N33/574G01N33/577A61K39/395A61K39/44
Inventor 宋怀东孔令春於惠敏
Owner RUIJIN HOSPITAL ATTACHED TO SHANGHAI NO 2 MEDICALUNIV
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