The present invention provides novel antibody sequences that bind and neutralize hCMV, and that can be used for preparing compositions for detecting, treating, inhibiting, preventing, and/or ameliorating hCMV infection or an hCMV-related disease. A population of immortalized, human B cells was divided in subcultures, and each subculture was tested for the presence of antibodies in the cell culture supernatant that bind and neutralize hCMV. Among the neutralizing subcultures, the isotype and clonality was determined for the antibodies secreted by the subculture named 1F7. These antibodies recognize a segment in the hCMV envelope glycoprotein H (gH) known to be bound by antibodies that neutralize hCMV infection. The antibody secreted by this subculture has been purified and the neutralizing ability confirmed using in vitro models for hCMV infection. The DNA sequences that encode the variable regions of the antibody secreted by the 1F7 subculture were amplified, cloned, and sequenced. The corresponding protein sequences were analyzed to identify the Complementarity Determining Regions (CDRs) that are responsible for the hCMV-specific biological activity. These sequences can be used for producing recombinant proteins having hCMV-specific binding and neutralizing properties, in the form of full antibodies, antibody fragments, or any other format of functional protein (e.g. bioactive peptide, fusion proteins) using appropriate expression vectors, host cells, and protein purification technologies. Compositions having therapeutic, prophylactic, and/or diagnostic utility in the management of hCMV infection and hCMV-related disorders can be prepared using these recombinant proteins, or the antibodies purified from cell cultures that have been generated using the 1F7 subculture
Antibodies against Human Cytomegalovirus (HCMV)
FUNARO, Ada;GRIBAUDO, Giorgio;LANDOLFO, Santo Giuseppe
2009-01-01
Abstract
The present invention provides novel antibody sequences that bind and neutralize hCMV, and that can be used for preparing compositions for detecting, treating, inhibiting, preventing, and/or ameliorating hCMV infection or an hCMV-related disease. A population of immortalized, human B cells was divided in subcultures, and each subculture was tested for the presence of antibodies in the cell culture supernatant that bind and neutralize hCMV. Among the neutralizing subcultures, the isotype and clonality was determined for the antibodies secreted by the subculture named 1F7. These antibodies recognize a segment in the hCMV envelope glycoprotein H (gH) known to be bound by antibodies that neutralize hCMV infection. The antibody secreted by this subculture has been purified and the neutralizing ability confirmed using in vitro models for hCMV infection. The DNA sequences that encode the variable regions of the antibody secreted by the 1F7 subculture were amplified, cloned, and sequenced. The corresponding protein sequences were analyzed to identify the Complementarity Determining Regions (CDRs) that are responsible for the hCMV-specific biological activity. These sequences can be used for producing recombinant proteins having hCMV-specific binding and neutralizing properties, in the form of full antibodies, antibody fragments, or any other format of functional protein (e.g. bioactive peptide, fusion proteins) using appropriate expression vectors, host cells, and protein purification technologies. Compositions having therapeutic, prophylactic, and/or diagnostic utility in the management of hCMV infection and hCMV-related disorders can be prepared using these recombinant proteins, or the antibodies purified from cell cultures that have been generated using the 1F7 subcultureI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.