Patent ID: 8916504

Claim:
A method of screening an scFv antibody RNA display library, the method comprising the steps of: (a) providing a library of puromycin or analogue thereof crosslinked single chain Fv (scFv) mRNA molecules, said molecules comprising an mRNA encoding a 5′ scFv and a 3′ spacer sequence, which molecules are crosslinked to a single stranded nucleic acid linker, the linker comprising a puromycin, or analogue thereof, at a 3′ end and a Psoralen C6 at a 5′ end; (b) in vitro translating the puromycin-crosslinked scFv mRNAs in the presence of a label under mildly oxidizing conditions using oxidized glutathione/reduced glutathione such that labeled puromycin-crosslinked scFv mRNA/protein molecules are formed; (c) purifying the labeled puromycin-crosslinked scFv mRNA/protein molecules; (d) screening the purified labeled puromycin-crosslinked scFv mRNA/protein molecules with at least one antigen; and (e) recovering the purified labeled puromycin-crosslinked scFv mRNA/protein molecules using magnetic beads; and optionally (f) reverse transcribing the scFv mRNA after antigen selection to make a cDNA that is complementary to the scFV mRNA.