humanizing an antibody? - (Jul/17/2007 )

hey there!

can anyone give me a good jumping-off point to learn how to do this? I have only the vaguest idea and I'd like to find out a bit more detail. I don't need a specific protocol at this point, but does anyone have any ideas on the best source of information on this topic?

thank you

A

check this from wikipedia:

humanized antibodies

hope it helps

good god, wikipedia and google really do know everything

thank you, mdfenko, but I was hoping for something a bit more specific...

I know that there are two methods:
1. make an immunoglobulin knockout mouse. add human immunoglobulin genes and get them to express. immunize your mouse with your chosen antigen. purify antibodies
or
2. clone the antibody entirely (cut and paste the dna, transfect it into cells for expression, etc) and try to find cells that will produce it, then make a hybridoma

but, I need something a bit more in the way of detail. from the time I have spent on google, pubmed, etc....it seems that several years ago (late 90's) there were a few labs that were working on this sort of tech. now, suddenly, nobody's publishing anything but you can find links to sites where it is possible to contract out the work. I also have tried to find immunoglobulin knockout mice to save some steps, or even human Ig-producing mice (we're not a poor lab) to save a lot of time, but have so far been unsuccessful.

I suspect either method above will take a long time - best-case scenario a couple of years, worst-case I tear my hair out in frustration 6 years from now when I can't get the protein to express, and some other lab beats me to it...

I was hoping someone here had actually done it, but it seems to be more an 'industry' sort of thing and most of the people here are academics.

Do you need total humanisation? I heard that you can graft the CDR into human Ig gene constructs.

well, yeah...the patients sometimes have poor reactions to the mouse sequences, and it's a real pain to write up a clinical trial when you have to include all that stuff too.