RE: Have it !! More Affimers28 Jun 2022 15:29
After the third pan, individual clones were screened by phage ELISA, and hits were classified as wells with a more than 2-fold increase in signal relative to controls (TcdA for TcdB and cell lysate for GDH). All hits were sequenced, and unique Affimer reagents were produced and purified. Binding to TcdB or GDH was assessed by ELISA (Figures S2A and S3A). Affimers that showed the highest signal by ELISA were taken forward, and pairwise binding of other Affimers was assessed by sandwich ELISA (Figures S2B and S3B). Affimers 18 and 45 were identified as the best pair to bind distinct sites of TcdB. No binding was observed in negative controls or against TcdA. GDH is hexameric, and Affimer 4 was demonstrated as the best capture and detection reagent.