Construction method of T-cell tumor phage antibody library and screened monoclonal antibody

Abstract

The invention belongs to the field of gene engineering, and particularly relates to a construction method of a T-cell tumor phage antibody library. The method comprises the steps of amplifying heavy chains and heavy chain variable genes of a full set of human antibodies from peripheral blood mononuclear cells of a healthy crowd and T cell leukemia / lymphoma patients, crossing and overlapping to form an antibody SCFV (Single Chain Fragment Variable) gene segment, then inserting into a phage vector pCanTab5F, and constructing the T-cell tumor phage antibody library. The invention relates to a monoclonal antibody screened through the method. The special phage antibody library is constructed aiming at T-cell tumors, and has higher sensibility and specificity than an existing commercialized antibody library. The constructed T cell leukemia / lymphoma phage antibody library has the capacity being 2.70*10<8> and meets the standards of a large-capacity antibody library. The antibody library is mainly used for high-throughput screening anti-CDR3 high-affinity humanized antibodies targeting T cell leukemia / lymphoma.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a method for constructing a T-line tumor phage antibody library and a screened monoclonal antibody. Background technique [0002] T cell neoplasms (T Cell Malignances) are a class of malignancies originating from a single T lymphoid precursor cell, with unique histopathological and clinical features, accounting for 25% of all lymphoid system malignancies, including T lymphocytic leukemia / lymphoma . They are more aggressive and less responsive to conventional chemotherapy than B-cell leukemia / lymphoma. Currently, the main methods for treating T-cell leukemia / lymphoma are chemotherapy and allogeneic hematopoietic stem cell transplantation (Allo-HCT). Other methods include targeted therapy, radiotherapy, and immunotherapy. One of the main causes of leukemia relapse after radiotherapy, chemotherapy, and hematopoietic stem cell transplantation (HCT) is the occurrence o...

AI Technical Summary

Problems solved by technology

The traditional TCR CDR3 receptor library sequencing method is far from meeting the detection requirements of the huge diversity of the immune repertoire. The next-generation sequencing technology can realize the throughput of a single determination of hundreds of thousands to several million sequences, providing a new way for the immune repertoire Comprehensive and systematic research provides an efficient technical platform, making it possible for deep sequencing of the TCR CDR3 region to accurately assess the immune status of patients

TCRs from healthy volunteers are rich in diversity, and the diversity of TCRs in tumor patients is oligoclonal. In the study of T-lineage tumors, we also found that there are obvious defects in the diversity of immune repertoire in such patients, mostly in the form of oligoclonal status

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