| 摘要 |
PURPOSE:To obtain a human monoclonal antibody useful for diagnosing or preventing infectious diseases, by manifesting a gene of multiform human antibody from a cDNA through cloning by using primers for gene cloning for the L-and H-strands of a human monoclonal antibody. CONSTITUTION:Lymphocytes are separated from the blood of bone marrow- transplanted patient(s) with high antibody titer to human cytomegalovirus (hCMV) and made immortal, and a mRNA is then separated from the resultant lymphocytes, and a cDNA is synthesized by using the mRNA. Next, the cDNA is subjected to cloning by using H-chain gene cloning primers for a human monoclonal antibody composed of the sense primer corresponding to the 5'-terminal of formula I, etc., primer for L-strand gene cloning for a human monoclonal antibody consisting of the anti-sense primer corresponding to the 3'-terminal of formula II, etc., and the sense primer corresponding to the 5'-terminal of formula III, etc., and the anti-sense primer corresponding to the 3'-terminal of formula IV; and the DNA produced is incorporated into a manifestation vector to effect manifestation, thus obtaining the objective anti-hCMV human monoclonal antibody. |
