Construction of scFv Fragments from Hybridoma or Spleen Cells by PCR Assembly

Today, antibodies can be obtained from naive repertoires (Winter et al. 1994; Vaughan et al. 1996) or libraries of fully synthetic genes (Knappik et al. 2000), and in the last decade, numerous libraries have been described (reviewed in Mondon et al. 2008). Nonetheless, hybridomas have remained the predominant source of antibodies, and a wealth of well characterized and even unique clones exist and are continuing to be generated. There is, thus, great interest in immortalizing these clones, in the extreme case, as a computer file of the sequences, as well as in accessing the antibody in a variety of new formats. To obtain enough material