Anti-GRPR antibody, method for producing same, detection method, use of the antibody, kit and gene construct

A gastrin-releasing peptide and antibody technology, applied in the fields of medicine, immunology and molecular biology, can solve the problem of not discovering the invention and making suggestions or expectations, and achieve the effect of auxiliary diagnosis

Inactive Publication Date: 2016-11-16
ZIEL BIOSCIENCIAS PESQUISA DESENVOLVIMENTO E DIAGNOSTICO LTDA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0025] Through a search of the literature, no literature was found that suggested or desired an existing invention

Method used

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  • Anti-GRPR antibody, method for producing same, detection method, use of the antibody, kit and gene construct
  • Anti-GRPR antibody, method for producing same, detection method, use of the antibody, kit and gene construct
  • Anti-GRPR antibody, method for producing same, detection method, use of the antibody, kit and gene construct

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0129] Example 1. Development and acquisition of antigenic peptides

[0130] Most immunogenic sequences of the gastrin releasing peptide receptor were identified using an antigenic index. It is an algorithm commonly used to create linear surface profiles of proteins. It is mainly obtained by computer programs to generate sets of values ​​and combine these sets of values ​​with values ​​​​predicted from the flexibility and secondary structure regions of polymer chains. This is because most of the antigenic domains are not only present in the surface contact regions of the protein. Therefore, it can be used to identify the second extracellular loop as the most immunogenic followed by ESTNQTFISCAPYPHSN (Glu-Ser-Thr-Asn-Gln-Thr-Phe-Ile-Ser-Cys-Ala-Pro-Tyr-Pro-His -Ser-Asn), this can be found in Figure 11a and Figure 11b see in. The main difference of the natural peptide of the gastrin releasing peptide receptor is that the number of amino acids of the natural peptide is re...

example 2

[0138] Example 2. Development and acquisition of antibodies .

[0139] Synthetic peptides were generated following the sequence ESTNQTFISCAPYPHSN and carried polyclonal antibodies developed by rabbit immunization.

[0140] On the first day, rabbits were inoculated subcutaneously with 200 ug of antigen emulsified into an equal volume of oil emulsion. Two more inoculations with the above-mentioned antigen emulsified into an oil emulsion were carried out at ten-day intervals. Five days after the last vaccination, plasma was collected and tested for specific antibodies to the corresponding antigens using dot blot.

[0141] dot blot

[0142] For immunodetection and quantification of antibodies by dot blot, proteins (approximately 0.5 ug) were added to 0.45 micron nitrocellulose membranes. After drying at room temperature for 30 minutes, the nitrocellulose membrane was blocked with a blocking agent for 1 hour and then incubated with serially diluted serum diluted to 5% for blot...

example 3

[0143] Example 3. Development of reagents and characterization and (or) determination of dysplasia and (or) cervical neoplastic lesions-immunohistochemistry method of quantitative detection.

[0144] After obtaining rabbit polyclonal antibodies, laboratory testing is achieved using immunohistochemical techniques. Paraffin blocks of cervical tissue and tumor and non-tumor tissues of the cervix were selected for immunohistochemical detection. All samples were detected with the antibody of the present invention and a polyclonal anti-rabbit gastrin-releasing peptide receptor control antibody (Affinity BioReagents, Golden, CO, USA, Cat. No.: OPA1-15619) for experimental use only.

[0145] Briefly, after deparaffinization, inactivation of endogenous peroxidases, and cross-reaction with conventional blocking serum, 4 μM sections and diluted primary antibodies (1:200 antibodies in the present invention and 1: 50 control antibody) were incubated overnight at 4°C. Localization of ...

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Abstract

The present invention relates to an anti-GRPR antibody obtained by a method that includes the step of exposing at least one animal to a peptide with at least 60% identity with the sequence of SEQ ID NO: 1. The present invention also describes a method for producing the anti-GRPR antibody, a method for detecting uterine dysplastic and / or neoplastic lesions in vitro, the use of the anti-GRPR antibody, a kit for qualitative and / or quantitative detection of uterine dysplastic and / or neoplastic lesions, and a gene construct for expressing or producing the antigenic peptide. The present invention pertains to the fields of medicine, immunology and molecular biology.

Description

technical field [0001] The invention is useful in the fields of medicine, immunology and molecular biology. More specifically stated, the present invention relates to anti-gastrin releasing peptide receptor (ANTI-GRPR) antibodies obtained by contacting at least one animal with a peptide up to 60% identical to the sequence SEQ ID NO:01. The present invention also describes: the process of obtaining anti-gastrin-releasing peptide receptor (ANTI-GRPR) antibody; the in vitro detection method of dysplasia and (or) uterine tumor lesion; anti-gastrin-releasing peptide receptor (ANTI-GRPR) ) the use of antibodies; quantitative detection and (or) quantitative detection reagents for dysplasia tumors and (or) uterine lesions; the expression or production of antigenic peptides is described by genetic construction. technical background [0002] Cervical cancer is the second most common cancer in women in the world, with nearly 500,000 new cases each year. Cervical cancer incidence is e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/30G01N33/53C07K7/08
CPCC07K16/30G01N33/57442C07K16/286C07K2317/34C07K16/28C07K16/3069G01N33/57411
Inventor D·B·考内里奥C·B·德法利亚斯
Owner ZIEL BIOSCIENCIAS PESQUISA DESENVOLVIMENTO E DIAGNOSTICO LTDA
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