The heavy chain Fd genes and K chain genes of immunoglobulin were amplified by RT-PCR from PBL of three volunteer donors with HIV-positive. Phage antibody library was constructed with the Fd genes and K chain genes us...The heavy chain Fd genes and K chain genes of immunoglobulin were amplified by RT-PCR from PBL of three volunteer donors with HIV-positive. Phage antibody library was constructed with the Fd genes and K chain genes using pComb3 as vector. Three-round selection against coated gpl20 showed specific enrichment of phage antibodies. After the third round selection, 40 out of 50 clones exhibited gpl20 binding capacity. The specificity of the clones was verified by ELISA and competition inhibition ELISA. The V_H was derived from subgroups V_Ⅱ and V_Ⅲ, V_L belonged to subgroups V_K Ⅰ and V_K Ⅲ with DNA sequencing. These results suggest that the antibodies obtained are specific to gpi20.展开更多
文摘The heavy chain Fd genes and K chain genes of immunoglobulin were amplified by RT-PCR from PBL of three volunteer donors with HIV-positive. Phage antibody library was constructed with the Fd genes and K chain genes using pComb3 as vector. Three-round selection against coated gpl20 showed specific enrichment of phage antibodies. After the third round selection, 40 out of 50 clones exhibited gpl20 binding capacity. The specificity of the clones was verified by ELISA and competition inhibition ELISA. The V_H was derived from subgroups V_Ⅱ and V_Ⅲ, V_L belonged to subgroups V_K Ⅰ and V_K Ⅲ with DNA sequencing. These results suggest that the antibodies obtained are specific to gpi20.
