Phage — Display Libraries of Murine Antibody Fab Fragments
作者： Jan Engberg , Liselotte Brix Jensen , Ali Fazil Yenidunya , Katja Brandt , Erik Riise
DOI: 10.1007/978-3-662-04605-0_5
关键词:
摘要: In this chapter we describe efficient procedures for construction, expression and screening of comprehensive libraries murine antibody Fab fragments displayed on the surface filamentous phage. Phagemid vectors are used placing randomly paired light (L) heavy (H) chain coding regions under transcriptional control Plac. The L (or H) region is fused in-frame with phage gene, ΔgIII, a truncated version protein pIII (ΔpIII). After superinfection helper induction P lac, Fd (composed VH CH1 domains) x chains assemble into in periplasm, Fab-ΔpIII complex at one end by displacing more) wild-type proteins. Enrichment phages affinity specific antigen then carried out successive rounds purification using antigen-coated microtiter wells, immunotubes or plastic beads followed reinfection E. coli cells eluted bound (1-6). An outline method illustrated Figure 1.
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