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Polypeptides capable of binding to CD19 and applications thereof

A nanobody and sequence technology, which is applied in the field of biomedicine, can solve the problems such as the listing of cell therapy drugs for B-cell malignant tumors.

Active Publication Date: 2021-08-31
Y CLONE MEDICAL SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are some CD19-CART cell therapy drugs in the U.S. market, but there is no cell therapy drug for B cell malignancies on the market in China. Therefore, it is urgent to develop antibodies against CD19 for the preparation of B cell malignancies and other CD19-related drugs. medicines for the treatment of diseases

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  • Polypeptides capable of binding to CD19 and applications thereof
  • Polypeptides capable of binding to CD19 and applications thereof
  • Polypeptides capable of binding to CD19 and applications thereof

Examples

Experimental program
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Effect test

Embodiment Construction

[0042] 1. Preparation of Immunogen

[0043] Based on the protein sequence and gene sequence information of human CD19 on the NCBI website, we analyzed and designed a polypeptide sCD19 that can effectively induce alpacas to produce specific antibodies against human CD19 protein, and connect His-tag (sCD19-his) or Rabbit Fc (sCD19-rFc) was used for subsequent purification and detection.

[0044] 2. Alpaca immunity and antiserum acquisition

[0045] The alpaca was primed with an emulsified mixture of 250 μg sCD19-rFc protein and 250 μl Freund’s complete adjuvant, and boosted three times with sCD19-rFc protein and 250 μl Freund’s incomplete adjuvant on the 14th day, 28th day, and 42th day 1 week after the third and fourth 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.

[0046] The titer of antiserum was detected by ELISA, and the detection pl...

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Abstract

The present invention relates to a polypeptide that can bind to human CD19. The present invention is aimed at the development of nanobody drugs and diagnostic kits for B cell malignant tumors. Through the preparation of CD19 protein, immunization of alpacas, and the use of phage library to display nanometer monoclonal antibody platform technology, etc., the nanobody VHH that specifically binds to CD19 is screened, The CDR sequence was identified, and the humanized antibody C19NB was constructed; at the same time, the binding of the humanized antibody to the CD19 protein on the cell surface was identified by flow cytometry. The invention provides a potential nanobody new drug for the clinical treatment of cancers expressed by CD19 antibodies, such as B cell 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 CD19, and the application of the polypeptide in drugs for treating malignant tumors of B cells. Background technique [0002] B-cell malignancies are common hematological malignancies. Although most patients respond to the current first-line treatment, the recurrence rate is high and the prognosis is poor. B-cell malignancies are still one of the refractory malignancies. In recent years, CD19 has received great attention as a molecular target for immunotherapy of B cell malignancies. [0003] CD19 is a specific surface protein of normal and malignant B lymphocytes. It is hardly expressed on the surface of other cells. It participates in the regulation of the PI3 / AKT pathway and plays a vital role in the proliferation and differentiation of B cells. Due to the specificity of CD19 expression in B lymphocytes and the generality of exp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/28C12N5/10C07K16/46A61K39/395A61K48/00A61P35/00G01N33/577G01N33/574
CPCA61K2039/505A61P35/00A61K39/001112C07K16/2803C07K2317/24C07K2317/31C07K2317/565C07K2317/569C12N5/0636C12N2510/00G01N33/57407G01N33/57492G01N33/577
Inventor 吴喜林吴稚伟黄碧莲施海霞
Owner Y CLONE MEDICAL SCI CO LTD