Fig. 2: Optimization of Snoopligase reaction conditions and immobilization. | npj Vaccines

Fig. 2: Optimization of Snoopligase reaction conditions and immobilization.

From: Development of a broad-spectrum subunit vaccine against H9N2 avian influenza using HA stem domain scaffold and snoopligase system

Fig. 2

a pH optimization. b Reaction time optimization. c Glycine concentration optimization. d SnoopTagjr substrate concentration optimization. e DogTag substrate concentration optimization. f SnoopTagjr and DogTag dual substrate concentration optimization. g Snoopligase concentration optimization. h Schematic representation of CL6B epoxidation and protein immobilization. i Schematic diagram of immobilized Snoopligase in the ligation reaction and purification of products. j Snoopligase immobilization: 1–2 before and after immobilization. k Purification of reaction products after ligation: 1 SnoopTagjr-EGFP, 2 IMX-DogTag, 3 Snoopligase, 4 unbound enzymes and substrates reaction, 5 immobilized enzymes and substrates reaction pH 2.0, elution 6–7 unreacted substrates. Error bar, mean ± SD.

Back to article page