Construction And Expression Of A Murine/human Chimeric Antibody With Specificity For HFRSV

The hemorrhagic fever with renal syndrome is a kind of severe infectious diseases, threatening human health. There is no specific and efficient way to treat it up to now. Since the technology of McAb appeared, the use of it for clinic diagnosis, therapy and prevention had shown some encouraging results. However, because most McAb are of murine origin, it induces a human anti-mouse antibody response when being used in vivo. Human McAbs are difficult to obtain with an adequate combination of specificity and effector function. Recently protein engineering appears, it looks promising in converting mouse McAb into" human" McAb. In this study, total RNA was extracted from 3D8 hybridoma, which secreted McAbs with high hemagglutination inhibition activity and high neutralization activity against HFRSV. The mRNA of total RNA was transcripted reversely into cDNA with oligo (dT)₁₅ primers. The first strand cDNA was used as template, and amplified with general primers for mouse antibody by PCR. The products were isolated from the agarose gel electrophoresis and cloned into pUC18 vectors respectively. Then the nucleic acid sequence had been done by the Sanger's dideoxy-mediated chain-termination method. The results showed that the cloned genes encode VL or VH of mouse antibody. Between them, VL gene consisted of 330 bp encodes 112 aa, which derived possibly from mouse light chains (?) ; VH gene consisted of 360 bp encodes 118 aa, and belongs to mouse heavy chains subgroup (?) (A).