Nano antibody capable of being combined with human CD38 and application thereof

A monoclonal antibody, sequence technology, applied in the field of biomedicine

Active Publication Date: 2020-10-23
Y CLONE MEDICAL SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In July 2019, Johnson & Johnson’s world’s first CD38 monoclonal antibody, Daratumumab Injection, was launched in China for the monotherapy of adult patients with relapsed and refractory mul

Method used

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  • Nano antibody capable of being combined with human CD38 and application thereof
  • Nano antibody capable of being combined with human CD38 and application thereof
  • Nano antibody capable of being combined with human CD38 and application thereof

Examples

Experimental program
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Effect test

Embodiment Construction

[0072] 1. Preparation of Immunogen

[0073] Based on the CD38 protein sequence and gene sequence information on the NCBI website, we analyzed and designed a polypeptide sCD38 that can effectively induce alpacas to produce specific antibodies against the extracellular domain of CD38, and linked His-tag (sCD38-his) or rabbit Fc (sCD38-rFc) was used for subsequent purification and detection.

[0074] 2. Alpaca immunity and antiserum acquisition

[0075] The alpaca was primed with an emulsified mixture of 250 μg sCD38-rFc protein and 250 μl Freund’s complete adjuvant, and boosted three times with sCD38-rFc protein and 250 μl Freund’s incomplete adjuvant on the 14th day, 28th day, and 42th day 1 week after the second and third immunizations, blood was collected to detect the antiserum titer; 1 week after the fourth immunization, 100 ml of blood was collected for the construction of the phage antibody library.

[0076] The antiserum titer was detected by ELISA, and the detection p...

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Abstract

The invention relates to a polypeptide capable of being combined with human CD38. The polypeptide comprises three complementary determining regions CDR1-3, the CDR1 sequence is one of or comprises sequences shown in SEQ ID NO:1-36, the CDR2 sequence is one of or comprises sequences shown in SEQ ID NO:37-72, and the CDR3 sequence is one of or comprises sequences shown in SEQ ID NO:73-111. Accordingto the invention, nano antibody drug development and diagnostic reagent research and development are carried out on hematopoietic malignant tumors. By preparing CD38 protein and immunizing alpaca andby a platform technology for displaying a nano monoclonal antibody by utilizing a phage library and the like, the invention screens a nano antibody VHH which is specifically combined with CD38, identifies a CDR sequence of the antibody, and constructs a humanized antibody C38NB; and meanwhile, the curative effect of the C38NB in treating multiple myeloma is evaluated in vivo by utilizing a humanized mouse model. The invention provides a potential nano-antibody new drug and a cell therapy new drug for clinical treatment of hematopoietic malignant tumors, and also provides a corresponding detection reagent for diagnosis of hematopoietic malignant tumors.

Description

technical field [0001] The invention relates to the field of biomedicine. More particularly, it relates to a polypeptide that can bind to human CD38, and the application of the polypeptide in the preparation of CD38 detection agent or hematopoietic malignant tumor treatment drug. Background technique [0002] CD38, also known as cyclic ADP ribose hydrolase, is a transmembrane glycoprotein. CD38 is expressed very low on normal lymphocytes and myeloid cells, as well as some tissues of non-hematopoietic origin, while upregulation is found in many hematopoietic malignancies (such as multiple myeloma cells) and cell lines derived from hematopoietic malignancies . Therefore, for such CD38-positive tumor conditions, anti-CD38 antibodies can be used as detection agents or therapeutic agents. [0003] In general, anti-CD38 monoclonal antibodies can clear tumor cells with high expression of CD38 through antigen-antibody immune response mechanism, the main mechanism is through 1) di...

Claims

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Application Information

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IPC IPC(8): C07K16/28C07K16/40C07K16/00C12N15/13G01N33/574G01N33/577A61K39/395A61P35/00
CPCC07K16/2896C07K16/40C07K16/005G01N33/57407G01N33/57492G01N33/577A61P35/00C07K2317/22C07K2317/24C07K2317/565C07K2317/567C07K2317/569C07K2317/92G01N2333/70596A61K2039/505
Inventor 吴稚伟吴喜林黄碧莲
Owner Y CLONE MEDICAL SCI CO LTD
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