Preparation method of monoclonal antibody based on Raman spectroscopy and micro-droplet technology

Abstract

The invention discloses a preparation method of a monoclonal antibody based on Raman spectroscopy and a microdroplet technology. The method comprises the steps of creating a micro-fluidic control platform and a corresponding detection probe and detecting the antibody content in microdroplets containing cells at high flux, wherein the microdroplets contain hybridoma cells formed by fusion of post-immune cells of an antibody immune animal or immune cells and myelomas. The ultra-efficient new technical method is provided for the preparation of the monoclonal antibody, the monoclonal screening cycle is shortened greatly, and the antibody screening capability is improved by geometric times.

Description

technical field [0001] The invention relates to a technical method for cell sorting and identification for monoclonal antibody preparation. Background technique [0002] Monoclonal antibody technology is an important tool for modern life science and medical research and treatment. In 1975, Koehler and Milstein created the in vitro hybridoma technology and obtained mouse-derived monoclonal antibodies. This technology won the Nobel Prize in Medicine and Physiology in 1984 prize. In 1986, the US FDA approved the marketing of the first monoclonal antibody drug, almost 30 years ago. At present, more than 40 therapeutic antibody drugs have been approved for marketing around the world, achieving sales of more than tens of billions of dollars each year. There has been an upsurge in the development of antibody drugs both internationally and domestically. [0003] The reality of the coexistence of huge market prospects and existing technical problems and barriers will inevitably tr...

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Problems solved by technology

The second screening process is extremely complicated and requires a lot of manpower to complete the isolation of subcellular clones

[0019] 5. Due to the use of pressure on individual cells, shear forces will reduce cell survival

Therefore, the low survival rate has also become a major flaw of this technology

[0020] 5. Flow screening needs to fully clean and disinfect the pipeline system to ensure that the cells will not be polluted, and the risk of contamination is high

[0023] In view of the increasing demand for the wide application of monoclonal antibodies, the operation of traditional hybridoma methods is very complicated, especially the process of monoclonal selection, which not only takes a lot of time, but also consumes a lot of manpower, and the operator The proficiency of the method will also affect the results, so the success or failure of the experiment often depends on this step. Even if the fluorescent technology is combined with the robotic arm picking, it is still not a technology to directly detect the amount of antibody expressed by a single cell. The technical complexity is too high and the time-sensitive sex is low

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