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Phage antibody library

The phage antibody library technology emerged in the late 1980s, a new antibody preparation, antibody genes can be easily obtained. In autoimmune diseases, cancer and other diseases of the immune antibody library has been applied and achieved satisfactory results. Storage capacity and the diversity of antibodies in the antibody library is screened by the specific antibodies. Capacity the greater the richer the diversity of antibody libraries, from the possibility of antibody screening to the greater. Antibody library build process, most researchers have sought to build the capacity of the antibody library, but less attention on the diversity of the antibody library. The study were collected from 25 patients with 50 mL of peripheral venous blood in patients with mixed peripheral blood lymphocytes were separated and extracted total RNA, and the application of the primer has been proven to be amplified by all antibody variable region fragments, to ensure the diversity of antibody libraries sex. The PCR method selected and the set of conditions is whether the amplification of antibody variable region gene sequences is another important factor. The experiments using semi-nested PCR amplification. Using semi-nested PCR amplification in the second round of the introduction of the corresponding restriction sites in preparation for the next cloning. Primers used by the author more, if all the direct PCR requires a lot of samples, the use of semi-nested PCR can take advantage of the product in the first round as the second round of PCR template, and significant savings in the use of cDNA . Of references in the experiment to introduce four restriction sites, and almost not in the corresponding light and heavy chain variable region appears to avoid the abnormal recombinants in the digestion process, to ensure the accuracy and specificity of the experimental results. In summary, the study was successfully constructed pneumoconiosis phage antibody library, and storage capacity and diversity of the antibody library is guaranteed, and lay the foundation for for pneumoconiosis antibody screening, pneumoconiosis immunotherapy provides a new ideas.

26.7.12 09:31

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