humanized antibody - how to design primers for CDRs - humanized antibody (Jan/03/2007 )

hello, i would like to ask you if there is anybody who is experienced with preparing humanized antibody. i did not understand from the papers i have read the way how to found out the sequence of CDRs of monoclonal antibody and than the way how to do the crossing over between the human and mouse variable domains. thank you very much for your answers.
Zuzana

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Okay I don't have experience but do understand when read antibody humanization article's as I'm interested as well.

1. CDRs are franked by highly conserved regions and before one design primers one needs to analyse the sequence of the selected antibody which will enable one to design highly specific primers. Using literature reference you can identify CDRs by aligning your sequences against the previously published work. One key feature of CDRs is high variability of sequences in this region.

2. Mouse-to-Human: Delete the CDRs in human antibody gene sequence, and replace the human CDRs with the ones amplified above and test the newly constructed human antibody against the antigen of interest.

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