Method for rapidly acquiring variable region gene sequence of specific human antibody

Abstract

The invention which belongs to the technical field of molecular biology concretely relates to a method for rapidly acquiring a variable region gene sequence of a specific human antibody, and a method for preparing a fully human antibody. The method provided by the invention has simple, rapid and effective processes, because the entire process of separating CD19 positive lymphocyte to acquiring the variable region gene sequence of the specific antibody consumes no more than 14 days. The invention provides a good technical platform, which can be widely applied to variable regions of heavy chains and light chains of a core component of the specific fully humanized antibody of various epidemic / infective and tumorous diseases, and autoimmune diseases especially some emergent and serious pandemic diseases such as SARS, bird flu and the like, and provides an effective means for fully humanized antibody medicament production.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a method for quickly obtaining a specific human antibody variable region gene sequence and a method for preparing a fully human antibody. Background technique [0002] With the development of the economy and the improvement of the medical level, the overall prevalence of infectious and infectious diseases has declined, but some infectious or infectious diseases are still important factors that threaten human health. From the large-scale outbreak of SARS in 2002, to the hand, foot and mouth disease caused by the EV71 virus in 2008, to the global influenza pandemic caused by the H1N1 influenza virus in Mexico in 2009, due to the lack of effective prevention and control in a short time Once outbreaks occur, they quickly spread on a large scale or globally. Therefore, for the explosive and epidemic diseases caused by such viruses, it is very urgent to find effe...

AI Technical Summary

Problems solved by technology

Mouse-derived monoclonal antibody is a common monoclonal antibody drug prepared from mice, but it has obvious limitations: the mouse-derived monoclonal antibody has low binding ability to human complement components, and has weak affinity with Fc receptors on the surface of NK and other immune cells , the ADCC effect mediated is weak, the half-life in human blood circulation is short, mouse monoclonal antibody is immunogenic, and the host is prone to produce anti-antibodies to cause allergic reactions, which often cause adverse reactions in the human body, thus greatly restricting its use in clinical trials and the development of related application fields

Disadvantages: (1) Transgenes often have somatic mutations and other unique sequences, resulting in incomplete human sequences; (2) Since antibodies are assembled in mice, the resulting mAbs have a murine glycosylation pattern , so these mAbs are not fully human in the end; (3) Transgenic mice express less diversity of human Ig, and cannot produce IgG subclasses in the same mouse

[0013] However, in the above-mentioned fully humanized antibody technology, the preparation and production process is complex, and it takes a long time and cycle, which is not suitable for the development and production of antibody drugs for some sudden and epidemic diseases, and rapid mutation of certain pathogens or antigens seem powerless

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