Biotin-avidin-lentiviral expression vector and CAR-T (chemric antigen receptor T) cell preparation method and visualization scheme

Abstract

The invention provides a biotin-avidin-lentiviral expression vector and a CAR-T (chemric antigen receptor T) cell preparation method, and aims at solving the defects of single targeting ability, complex preparation, time-consuming and labor-consuming properties and poor economical efficiency of traditional CAR-T. The method comprises the following steps of constructing an avidin-CAR lentivirus package carrier; performing virus packaging; and performing T cell separation, activation, co-stimulation structural domain gene transduction and amplification, thereby preparing CAR-T cells. On the basis of a general CAR-T treatment system, a molecular imaging detection means is introduced, a cell-based visualization treatment scheme is provided, and a CAR-T cell treatment process can be dynamically monitored in vivo in real time, so that a situation that the current CAR-T cell treatment process is lack of effective monitoring is solved. The biotin-avidin-lentiviral expression vector has the advantages of simplicity in preparation, diversity in selection, extensive targets and visualization on treatment process, and suitability for treatment of tumor patients.

Description

technical field [0001] The invention belongs to the field of biomedicine, and mainly relates to preparation of chimeric antigen receptor T cells and visualization during treatment. Especially a biotin-avidin-lentiviral expression vector and CAR-T cell preparation method and visualization scheme. Background technique [0002] Biotin-avidin system (Biotin-Avidin-System, BAS) is a new biological reaction amplification system developed in the late 1970s. With the advent of various biotin derivatives, BAS was soon widely used in various fields of medicine. A large number of studies in recent years have confirmed that the biotin-avidin system can be combined with almost all kinds of markers that have been successfully studied so far. The strong combination of high affinity between biotin and avidin and the multi-level amplification effect make BAS immunolabeling and related tracer analysis more sensitive. It has become a new technology that is widely used in the qualitative and...

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to address the shortcomings of the existing traditional CAR-T treatment system, and to solve the single targeting, cumbersome preparation, and time-consuming problems of traditional CAR-T by preparing a new and universal CAR-T treatment system. The shortcomings of labor and poor economy; and on the basis of the prepared general-purpose CAR-T treatment system, molecular imaging detection methods are introduced to monitor the CAR-T treatment process in real time, dynamically and in vivo, and the general-purpose CAR-T treatment system can be visualized. So as to solve the current situation that the current CAR-T system lacks effective monitoring during the treatment process

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