Method and system for screening specific BCR/TCR

Abstract

The invention discloses a method and a system for screening specific BCR / TCR. The method for screening the specific BCR / TCR comprises the following steps of (1) extracting mRNA in lymphocytes of an immunized sample to be detected; (2) carrying out reverse transcription treatment on the mRNA obtained in the step (1), then carrying out multiple PCR amplification on the BCR / TCR variable region gene, then carrying out high-throughput sequencing, and determining nucleic acid and amino acid sequences of the BCR / TCR variable region; and (3) comparing the sequence obtained by sequencing with a lymphocyte BCR / TCR sequencing result of a sample which is not stimulated by the antigen to obtain a corresponding specific immune sequence. The method disclosed by the invention has the advantages that by comparing big data of the immune spectrum and comparing a huge number of BCR / TCR sequences, the specificity and the accuracy are higher, the efficiency of screening and identifying the antibody and the T cell receptor can be greatly improved, and the screening cost is low.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and cellular immunology, and in particular relates to a method and system for screening specific antibodies (BCR) and T cell receptors (TCR). Background technique [0002] Monoclonal antibody is a highly uniform antibody that recognizes a specific epitope produced by a single B cell. Monoclonal antibodies have broad application prospects in basic medical and biological research, as well as in the diagnosis, prevention and treatment of clinical diseases. To obtain monoclonal antibodies against specific antigens, antibody screening is required. The existing relatively mature monoclonal antibody screening technologies include: [0003] 1. Hybridoma technology. [0004] The mouse splenocytes after antigen immunization are fused with mouse myeloma cells, and the hybrid cells formed can not only produce antibodies, but also proliferate indefinitely. The fused hybrid cells are divided into...

AI Technical Summary

Problems solved by technology

[0011] The disadvantages of monoclonal technology are twofold: on the one hand, the number of BCR or TCR on the surface of a single lymphocyte is limited, which limits the sensitivity and specificity of single cell sorting by antigen, and the sorted cells cannot be guaranteed to be positive cells. In the later stage, more identification of the obtained antibody / TCR is still required; on the other hand, the difficulty of single cell cloning is relatively high, and there are high requirements on the quality of cells, the skills of operators, and the quality of reagents

[0012] There are still three significant deficiencies in the existing monoclonal antibody screening technology: the first is that it is difficult to screen human antibodies, because it is impossible to repeatedly immunize the human body with antigens and adjuvants like immunized animals

[0013] The second is the difficulty in preparing monoclonal antibodies against complex antigens

When it is necessary to prepare antibodies against larger and more complex antigens (such as membrane protein complexes, tumor cells, etc.), it is difficult to select and prepare antigens for immunization and there is a risk of failure, and the function cannot be guaranteed after immunization Antibodies that meet the above requirements (such as antibodies obtained by immunizing with polypeptide fragments, not necessarily neutralizing antibodies)

If multiple antigens are selected for immunization and screening at the same time, the cost of monoclonal antibody preparation will be significantly increased

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