| 摘要 |
Disclosed is a method of screening an scFv antibody RNA display library, the method comprising the steps of: (a) providing a puromycin or analogue thereof crosslinked single chain Fv (scFv) mRNA molecule, said molecule comprising an mRNA encoding a 5' scFv and a 3' spacer sequence, which molecule is 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 mRNA molecule in the presence of a label under mildly reducing conditions such that a labeled puromycin-crosslinked scFv mRNA/protein molecule is formed; (c) purifying the labeled puromycin-crosslinked scFv mRNA/protein molecule; (d) subjecting the purified labeled puromycin-crosslinked scFv mRNA/protein molecule to antigen selection with at least one antigen; and (e) recovering the purified labeled puromycin-crosslinked scFv mRNA/protein molecules using affinity based magnetic beads.
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