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Biotech / Medical : Biotech for less than cash value
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To: Hank who wrote (159)8/10/2003 12:12:44 PM
From: AuBug  Read Replies (1) of 684
 
The mAbs displayed on phage come from humans. E.g., you extract B cells from convalescent patients that survived a viral disease and build a cDNA library. You then insert the variable region of the human antibodies into a phagemid vector and express it in E coli. The reducing environment of the periplasmic space of the bacterium forms the inter- and intra- disulfide bonds so you get the same folding and conformations. You may not get the glycosylation but you can turn around and express them in yeast and get essentially the same as humans. The genes are 100% human. You then pan (screen) the phage display library against some antigen, e.g. the virus, and select those that bind it. You then test them for binding affinity and neutralization. You can also improve them by using error-probe PCR mutagenesis or CDR shuffling or other techniques to make numerous permutations and test them for improved performance. This scenario would be different if you were looking for mAbs against endogenous proteins such as receptors or transporters or phosphorylation sites or whatever.

My point is that phage display offers access to 100% hmAbs derived from the full range of possible VDJ combinations of the antibodies produced by humans. These huMouse technologies, which only use a subset of human genes, can and do produce some very high affinity mAbs to the desired therapeutic target. For my money phage display is superior but not the only functional approach.
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