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Scripps Research scientists devise powerful new method for finding therapeutic antibodies
Date:9/11/2012

ntibodies and inserting them into lentiviruses. Unlike the phage viruses used in traditional methods, lentiviruses can usefully infect mammalian cells, delivering their payloads antibodies, in this case into a more human-like cellular environment.

Zhang applied this new library of antibody-coding lentiviruses to a single, large culture of mammalian test cells. The cells were of a type that express EPO receptors and proliferate when these receptors are bound by EPO proteins or by antibodies that effectively mimic EPO. Each of these cells could host only a few viral particles at most, so in this way Zhang was able to distribute the entire library of EPO-receptor-binding antibodies broadly within the cell culture. Zhang also cultured the cells in a special way that prevented antibodies secreted by one cell from spreading easily to nearby cells and muddying any cause-effect relationship. "This concern over the diffusion of antibodies in the culture was one of the factors that had discouraged other researchers from using such a technique," said Zhang.

After the lentiviruses had delivered the antibodies to the cultured cells, Zhang was able to note which cells were proliferating the most signifying the presence of antibodies that mimic EPO. To identify the antibodies responsible, Zhang had only to harvest these faster-growing cells and sequence the antibody genes inside them.

This method quickly yielded an antibody that in a further test showed about 60 percent of the biological activity of natural EPO which was as good as any antibody EPO-mimic that had ever been described.

Opening the Door to the Unknown

But Zhang and Lerner also noted that many of the proliferating cells had been infected by multiple lentivirus particles, and contained sequences from more than one antibody. Puzzlingly, Zhang found that when he recreated antibodies from these sequences, and tested them individually or in combinations, they showed
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Contact: Mika Ono
mikaono@scripps.edu
858-784-2052
Scripps Research Institute
Source:Eurekalert  

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