Fig. 2.

Demonstration of target-specific ligand selection by using CIS display. (A) A 1:1 mixture of Cκ-RepA DNA and V5-repA DNA was prepared, transcribed and translated in vitro, and selected against either anti-Ck antibody, anti-V5 antibody, or anti-ACTH antibody. Recovered DNA was amplified with universal primers and separated by agarose gel electrophoresis on the basis of the size difference between the Cκ domain and the V5 peptide tag. The amount of DNA specifically recovered when by using the anti-Cκ and anti-V5 antibodies is shown in duplicate along with a single negative control (anti-ACTH) to reflect background recovery. (B) A 1:10⁸ dilution of Cκ-RepA DNA into V5-repA DNA was prepared and subjected to four rounds of selection against either the anti-Cκ antibody or the anti-FLAG antibody as a negative control. The four rounds of selection are indicated along with the positions of the two recovered PCR products. (C) A 1:10¹⁰ dilution of Cκ-RepA DNA into V5-repA DNA was prepared and subjected to four rounds of selection against either the anti-Cκ antibody or the anti-FLAG antibody as a negative control. The four rounds of selection are indicated along with the positions of the two recovered PCR products. Marker refers to the DNA Hyperladder (Bioline, London).