I am trying to generate monoclonal antibodies (murine Fab, BALB/c mice) by the phage display method. Many studies focused on amplifying the kappa region of the light chain. Isn't that both kappa chain and lambda chain are important to be included in the pool of light chain fragments? Thank you.
Is that still the case? We had to design our own primer sets more than 20 years ago ( https://plueckthun.bioc.uzh.ch/wp-content/uploads/Publications/APpub0113.pdf , updated and improved in https://plueckthun.bioc.uzh.ch/wp-content/uploads/Publications/APpub0307.pdf ), because the primer sets available at the time failed to amplify some of our most interesting antibodies - which turned out to be murine lambda chains.
Of course, the lab mice most frequently used for monoclonal antibody production use only one lambda VL segment, in contrast to wild mice. Murine lambda chains are tricky to humanize because of a particular substitution of a conserved VL/VH interface residue that can strongly affect antigen affinity by allowing more flexibility in the interface (https://plueckthun.bioc.uzh.ch/wp-content/uploads/Publications/APpub0178.pdf).
Back then, many were of the opinion that the murine lambda are not used at all,, but our experience proved that this is not the case.
Is that still the case? We had to design our own primer sets more than 20 years ago ( https://plueckthun.bioc.uzh.ch/wp-content/uploads/Publications/APpub0113.pdf , updated and improved in https://plueckthun.bioc.uzh.ch/wp-content/uploads/Publications/APpub0307.pdf ), because the primer sets available at the time failed to amplify some of our most interesting antibodies - which turned out to be murine lambda chains.
Of course, the lab mice most frequently used for monoclonal antibody production use only one lambda VL segment, in contrast to wild mice. Murine lambda chains are tricky to humanize because of a particular substitution of a conserved VL/VH interface residue that can strongly affect antigen affinity by allowing more flexibility in the interface (https://plueckthun.bioc.uzh.ch/wp-content/uploads/Publications/APpub0178.pdf).
Back then, many were of the opinion that the murine lambda are not used at all,, but our experience proved that this is not the case.
Perhaps the original reason was that the vast majority of of murine hybridomas produce antibodies comprising kappa light chains without any pre-selection, leading to the idea that this part of the repertoire is enough. But I agree that this should not be a fair conclusion. The experience with human libraries gave us a clear message. Some of the best binders (obtained by different researchers in different labs with independent libraries) are lamdbda-containing antibody fragments. In fact, they tend to be more secreted and/or stable as soluble fragments outside the phages than their kappa counterparts. There are not many studies with mouse lambda-containing antibody fragments, but that field deserves a better exploration for sure.
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