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Interleukin 2 binding molecule, and derivative, kit, production method and use of interleukin 2 binding molecule

A technology of interleukin and binding molecules, which is applied in the field of single-domain antibodies against interleukin 2, can solve the problem of fewer antibodies and achieve high affinity, high stability, and high specificity

Active Publication Date: 2021-08-13
XIAMEN BIOCHEE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although antibodies against IL2 have been developed, there is still a need for improved anti-IL2 antibodies as therapeutic agents
In addition, it is worth noting that there are currently few single domain antibodies against IL2

Method used

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  • Interleukin 2 binding molecule, and derivative, kit, production method and use of interleukin 2 binding molecule
  • Interleukin 2 binding molecule, and derivative, kit, production method and use of interleukin 2 binding molecule
  • Interleukin 2 binding molecule, and derivative, kit, production method and use of interleukin 2 binding molecule

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] This embodiment discloses an interleukin 2 binding molecule, which can specifically bind interleukin 2, and comprises at least one immunoglobulin single variable domain, and at least one immunoglobulin single variable domain comprises CDR1, CDR2 and CDR3. wherein CDR1 comprises an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 1-15, CDR2 comprises an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 16-54, and CDR3 comprises an amino acid sequence identical to any one of SEQ ID NOs: 16-54. An amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 55-84. In this embodiment at least one immunoglobulin single variable domain comprises CDR1, CDR2 and CDR3 shown in Table 1 below.

[0065] Table 1 CDR1, CDR2 and CDR3 of interleukin-2 binding molecules

[0066]

[0067]

[0068]

[0069]

[0070]

[0071] Among them, the immunoglobulin single variable domain is VHH. The VHH comprises amino acid se...

Embodiment 2

[0076] This example discloses the screening production process of Example 1, and the 54 antibody strains obtained through screening are subjected to nucleotide sequencing to obtain nucleic acid sequences.

[0077] 1. Material preparation: Prepare the commercial materials shown in Table 3 below.

[0078] table 3

[0079]

[0080] 2. Immunization

[0081] To induce a humoral immune response against IL2 in camelids, alpacas were injected subcutaneously with 5 doses of human IL2 protein at 1 to 3 week intervals. The dosage range is 150ug to 500ug per injection.

[0082] 3. Serum titer detection

[0083] After immunization, serum titers of anti-IL2 specific antibodies were determined by ELISA. For ELISA experiments, ELISA (NEST, 504201) was coated with 400 ng / well recombinant his-tagged human IL2 protein and incubated overnight at 4°C. After blocking and washing, serially diluted preimmune and immune sera were added and incubated at room temperature for 2 hours, then incuba...

Embodiment 3

[0114] This embodiment discloses an immunoconjugate and a pharmaceutical composition.

[0115] The immunoconjugate comprises a therapeutic moiety and the interleukin-2 binding molecule in Example 1. Therapeutic moieties include cytotoxins, biologically active proteins or radioisotopes. The therapeutic moiety is conjugated to the interleukin-2 binding molecule.

[0116] The pharmaceutical composition comprises the interleukin-2 binding molecule in Example 1 or the above immunoconjugate, and a pharmaceutically acceptable carrier.

[0117] The pharmaceutical composition is a drug for preventing or treating a proliferative disorder, a disease or condition that may respond to an IL2 antagonist: such as a drug for treating cancer, or a drug for treating chronic viral infection, or a drug for inhibiting or blocking the binding of CD25 and interleukin 2.

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PUM

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Abstract

The invention discloses an interleukin 2 binding molecule, which can specifically bind to interleukin 2 and comprises at least one immunoglobulin single variable domain; the at least one immunoglobulin single variable domain comprises CDR1, CDR2 and CDR3, wherein CDR1 comprises an amino acid sequence that is at least 90% identical to any one of SEQ ID NO: 1-15; CDR2 comprises an amino acid sequence that is at least 90% identical to any one of SEQ ID NO: 16-54; and CDR3 comprises an amino acid sequence that is at least 90% identical to any one of SEQ ID NO: 55-84. The invention also discloses a nucleic acid molecule encoded by the interleukin 2 binding molecule, an expression vector, a host cell, a production method, an immunoconjugate using the interleukin 2 binding molecule, a pharmaceutical composition, a kit and application of the interleukin 2 binding molecule. According to the invention, the interleukin 2 binding molecule with high specificity, high affinity and high stability is obtained through screening and is used for treating, preventing and diagnosing IL2 related diseases.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a single-domain antibody against interleukin 2 (IL2). Background technique [0002] Interleukin 2 (interleukin 2, IL2) has been discovered for more than 30 years, but it is still one of the most concerned and widely studied cytokines. IL2 in the human body is a glycoprotein with a molecular weight of 15.5kDa. The mature IL2 molecule consists of 133 amino acid residues left after the signal peptide with 20 amino acid residues is cut off from the N-terminus of the peptide chain of 153 amino acids. IL2 and other cytokines in No homology in sequence (T Taniguchi, H Matsui, T Fujita, C Takaoka, NKashima, R Yoshimoto, J Hamuro. Structure and expression of a cloned cDNA for human interleukin-2. Nature. 1983 Mar 24-30; 302(5906) :305-10.). In a steady state, IL2 is mainly produced by CD4+ T helper cells in secondary lymphoid organs, and a small part is produced by CD8+ T cells, NK ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/24C12N15/13A61K39/395A61K47/68A61K45/06A61P35/00A61P31/12
CPCC07K16/246A61K47/6845A61K45/06A61P35/00A61P31/12C07K2317/569C07K2317/565C07K2317/92C07K2317/94C07K2317/76C07K2317/22Y02A50/30
Inventor 梁耀极刘杰陈莹陈滨滨
Owner XIAMEN BIOCHEE BIOTECHNOLOGY CO LTD
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