Antibodies against PD-1 and uses therefor

A PD-1 and antibody technology, applied in the direction of antibodies, applications, anti-inflammatory agents, etc., can solve problems such as weakening of T cell response

Inactive Publication Date: 2010-12-01
WYETH LLC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conversely, engagement of the inhibitory CTLA-4 receptor with the same B7-1 or B7-2 ligand results in attenuation of the T cell response

Method used

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  • Antibodies against PD-1 and uses therefor
  • Antibodies against PD-1 and uses therefor
  • Antibodies against PD-1 and uses therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0130] Example 1: Selection of ScFv binding to PD-1

[0131] Using an expanded version of the 1.38 x 10 10 The library's scFv phagemid library selects antibodies specific for human PD-1. Soluble PD-1 fusion protein (20 μg / ml in phosphate-buffered saline (PBS) solution) or control fusion protein (50 μg / ml in PBS solution) was coated into microtiter plate wells and left overnight at 4°C. Wells were washed in PBS and blocked in MPBS (3% milk powder in PBS) at 37°C for 1 hour. The purified phage (10 12Transducing units (tu) were blocked for 1 hour in 3% MPBS in a final volume of 100 μl. Add blocked phage to blocked control fusion protein wells and incubate for 1 hour. Blocked and deselected phage were then transferred to blocked wells coated with PD-1 fusion protein and incubated for an additional hour. Wells were washed 5 times with PBST (0.1% v / v Tween 20 in PBS) and then 5 times with PBS. Bound phage particles were eluted and used to infect 10 ml of exponentially growin...

Embodiment 2

[0134] Example 2: Determination of antibody specificity to PD-1 by phage ELISA

[0135] To determine the specificity of the antibody against PD-1, phage ELISA was performed against PD-1 fusion protein and control protein. Individual E. coli colonies from selection products were picked into 96-well plates containing 100 [mu]l 2TYAG medium per well. M13K07 helper phage at a multiplicity of infection (moi) of 10 was added to the exponentially growing culture and the plate was incubated at 37°C for an additional 1 hour. The plate was centrifuged at 2000 rpm for 10 minutes in a table top centrifuge. The supernatant was removed and the cell pellet was resuspended in 100 μl 2TYAK and incubated overnight at 30°C with shaking. The next day, the plate was centrifuged at 2000 rpm for 10 minutes and the phage-containing supernatant from each well was transferred to a new 96-well plate. Phage samples were blocked in MPBS at a final concentration of 3% prior to ELISA.

[0136] 0.5-2.5...

Embodiment 3

[0138] Example 3: Identification of antibody clones

[0139] E. coli clones of PD-1 binding scFv were streaked onto 2TYAG plates and incubated overnight at 30°C. Amplification of V from scFv clones by using pCANTAB6 vector sequence oligonucleotides (oligos) H and V L Colonies from these plates were sequenced. Unique PD-1 binding clones that neutralized PD-L1 binding to PD-1 were assayed as described in Example 4. Sequence differences between scFv and IgG formats are due to changes introduced by PCR primers during conversion from scFv to IgG.

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Abstract

This disclosure provides antibodies and antigen-binding fragments that can act as agonists and/or antagonists of PD-1 (Programmed Death 1), thereby modulating immune responses in general, and those mediated by TcR and CD28, in particular. The disclosed compositions and methods may be used for example, in treating autoimmune diseases, inflammatory disorders, allergies, transplant rejection, cancer, and other immune system disorders.

Description

[0001] This application is a divisional application of the invention patent application titled "anti-PD-1 antibody and its use" with the international application PCT / IB2003 / 006304 entered into China with the international filing date on December 22, 2003 and the application number 200380109929.8. technical field [0002] The technical field relates to the modulation of immune responses regulated by the programmed death 1 (PD-1) receptor. Background technique [0003] The adaptive immune response involves the activation, selection and clonal expansion of two classes of lymphocytes called T cells and B cells. After encountering an antigen, T cells proliferate and differentiate into antigen-specific effector cells, while B cells proliferate and differentiate into antibody-secreting cells. [0004] T cell activation is a multistep process requiring several signaling events between T cells and antigen presenting cells (APCs). For T cell activation to occur, two types of signals...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C12N15/13C12N15/63C12N1/21C12N5/10A61K39/395A61P37/02A61P37/06A61P37/08A61P35/00
CPCC07K16/2803C07K2317/622A61K2039/505C07K16/2818C07K2316/96C07K2317/73C07K2317/76A61P1/04A61P19/02A61P25/00A61P29/00A61P31/00A61P35/00A61P37/02A61P37/04A61P37/06A61P37/08A61P43/00A61P3/10
Inventor M·科林斯C·伍德B·卡雷诺V·瓦尔格-阿切尔D·卢克森伯格J·朱西夫C·拉塞尔L·L·卡特F·本内特J·安德鲁斯
Owner WYETH LLC
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