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. Author manuscript; available in PMC: 2009 Sep 15.
Published in final edited form as: Clin Cancer Res. 2008 May 1;14(9):2647–2655. doi: 10.1158/1078-0432.CCR-07-1442

Figure 1. Method overview.

Figure 1

A-B, Enrichment of a yeast-display scFv library for scFv binding to markers by (A) 2 magnetic enrichments and (B) 3 fluorescent cell sortings. C, Depletion of the enriched library for cross-reactive scFv. D, PCR amplification of marker-specific recognition sequences and yeast co-transformation with the vector pTOR2 for cloning by gap repair. E, Growth and induction of yeast-secreting scFv in liquid medium. F, High-throughput Ni- purification of secreted scFv. G, Analysis for marker specificity by capture ELISA. H, Mating of yeast secreting marker-specific scFv with yeast producing biotin ligase. I, Growth and induction of diploid yeast to secrete marker-specific Bbs. J, large scale Ni-purification to obtain reagents for double determinant assays. K, Reagent ready for diagnostic tests such as bead-based assays.