A human scFv antibody generation pipeline for proteome research

摘要: The functional decryption of the human proteome is challenge which follows sequencing genome. Specific binders to every protein are key reagents for this purpose. In vitro antibody selection using phage display offers one possible solution that can meet demand 25,000 or more antibodies, but needs substantial standardisation and minimalisation. To evaluate potential, three human, naive gene libraries (HAL4/7/8) were constructed a standardised pipeline was set up. quality validated with 110 antigens, including other mammalian, fungal bacterial proteins, viruses haptens. Furthermore, abundance VH, kappa lambda subfamilies during library cloning E. coli based system on packaging scFvs evaluated from analysis 435 individual resulting in first comprehensive comparison V subfamily use all steps an pipeline. Further, compatible cassette vector mammalian expression fragments described, allowing vivo biotinylation, enzyme fusion Fc fusion.