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Novel pancortin-pablo protein interactions and methods of use thereof

A complementary sequence and polynucleotide technology, applied in the direction of anti-animal/human immunoglobulin, apoptosis-related protein, mammalian protein, etc., can solve the problems of loss of plasma membrane integrity, difficulty in apoptosis, cell swelling, etc.

Inactive Publication Date: 2005-08-17
WYETH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Apoptotic cells are rapidly and efficiently removed making detection of apoptosis in tissue sections extremely difficult
[0005] In contrast, necrosis is associated with rapid metabolic breakdown leading to cell swelling, early loss of plasma membrane integrity and eventual cell rupture

Method used

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  • Novel pancortin-pablo protein interactions and methods of use thereof
  • Novel pancortin-pablo protein interactions and methods of use thereof
  • Novel pancortin-pablo protein interactions and methods of use thereof

Examples

Experimental program
Comparison scheme
Effect test

specific example

[0124] A specific example of the production of the antibody of the present invention is provided below: about 1-200 μg of the antigen containing the polypeptide of the present invention is injected intraperitoneally into a mouse. The growth of B lymphocytes is stimulated by injection of the antigen and an adjuvant such as complete Freund's adjuvant (a non-specific stimulator of immune response, including inactivated Mycobacterium tuberculosis). After a period of time (for example, at least two weeks) after the first injection, mice are injected with a second dose of antigen mixed with incomplete Freund's adjuvant for booster immunization.

[0125] A few weeks after the second injection, blood was taken from the tail of the mouse, and the serum was titrated by immunoprecipitation against the radiolabeled antigen. It is best to repeat the booster immunization and titration process until a suitable titer is obtained. The mouse spleen with the highest titer was taken out, and the sple...

example

[0131] In addition, examples of methods and reagents particularly suitable for generating and screening antibody display libraries can be found in the following references: for example, U.S. Patent No. 5,223,409; International Application No. WO 92 / 18619; International Application No. WO 91 / 17271; International Application number WO92 / 20791; international application number WO 92 / 15679; international application number WO 93 / 01288; international application number WO 92 / 01047; international application number WO 92 / 09690 and international application number WO90 / 02809.

[0132] In addition, recombinant anti-pancortin antibodies and / or anti-pancortin-Pablo antibodies containing human and non-human fragments that can be prepared using standard recombinant DNA technology, such as chimeric antibodies and humanized monoclonal antibodies, are also within the scope of the present invention. The chimeric antibody and humanized monoclonal antibody can be prepared by recombinant DNA technol...

Embodiment 1

[0266] Identification of pancortin interacting protein

[0267] Yeast two-hybrid test

[0268]It was observed that pancortin 2 and pancortin 4 bind Pablo in the yeast two-hybrid test. Therefore, in the yeast two-hybrid test or the yeast three-hybrid test, pancortin protein, pancortin-Pablo dimer or part thereof can be used as a "bait protein" (see, for example, US Patent No. 5,283,317; and International Application No. WO 94 / 10300 ) To identify other proteins that bind or interact with pancortin and / or pancortin-Pablo and are involved in pancortin and / or pancortin-Pablo activity. The pancortin binding protein may also be involved in the signal transmission of the pancortin protein or pancortin target, for example as a downstream element of the Pablo-mediated signal pathway. Alternatively, the pancortin binding protein may be a pancortin inhibitor.

[0269] The two-hybrid system is based on the regulatory properties of most transcription factors consisting of independe...

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Abstract

This invention relates to newly identified human pancortin polypeptides, the interaction of the pancortin polypeptides with a Pablo polypeptide, the use of such polypeptides, as well as the production of such polypeptides. The invention relates also to identifying compounds which modulate the activity of a pancortin polypeptides and / or the interaction of a pancortin-Pablo polypeptide interaction, wherein modulators can be agonists, antagonists and / or inhibitors of pancortin and / or the pancortin / Pablo interaction and therefore potentially useful in therapy.

Description

Invention field [0001] The present invention generally relates to the fields of cell signaling, apoptosis, neuroscience and molecular biology. More specifically, the present invention relates to a newly identified polypeptide interaction comprising a neuron-specific pancortin polypeptide and a neuron-specific pro-apoptotic Pablo polypeptide, the application of the polypeptide, the regulation of the polypeptide, and the production of the polypeptide . The present invention also relates to the identification of compounds that can act as agonists, antagonists and / or inhibitors of pancortin-Pablo interaction, and therefore have potential therapeutic value. Background of the invention [0002] Apoptosis is a form of programmed cell death that occurs by activating the cell's intrinsic suicide mechanism. This biochemical mechanism responsible for apoptosis is expressed in most, if not all, cells. Apoptosis is mainly a physiological process necessary to remove individ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027A61K31/7088A61K38/00A61K45/00A61K48/00A61P9/10A61P17/06A61P35/00A61P43/00C07K14/47C07K16/18C07K19/00C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12P21/02C12P23/00C12Q1/02C12Q1/68
CPCA01K2217/075C07K14/4747A01K2217/05A61K38/00A61P9/10A61P17/06A61P35/00A61P43/00
Inventor R·J·马克A·T·伍德K·古卢科塔
Owner WYETH LLC
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