English: SPR analysis of the purified G6 scFv. A, Illustrative schematic representing of G6 scFv immobilization on an MUA‐modified Ausensor

slide at pH 4.5 by amine coupling protocol: (1) Formation of self‐assembled monolayers ofMUA. (2) Activation of carboxyl groups in MUA by EDC/NHS. (3) Immobilization of 250 μg/mL G6 scFv. (4) Deactivation of unreacted activated ester sites by ethanolamines. (5) Interaction of Hsp70 in the related peptide with G6 scFv. B, SPR sensogram of the G6 scFv immobilization on an MUA‐modified Au‐sensor slide that shows immobilization level and RU of the G6 scFv was approximately 3.6 times more than that of blank channel. C, Sensogram of Hsp70 in related peptide interaction with immobilized G6 scFv at pH 7.5 of PBS running buffer. Kinetic parameters after injection of different concentrations of Hsp70 in the related peptide (step 5 in Illustrative schematic) shown the G6 scFv had good affinity for Hsp70 in the related peptide (KD=7.22×10−7 M). EDC, N‐(3‐dimethylaminopropyl)‐N‐ethylcarbodiimide hypochloride; Hsp, heat shock protein; MUA, 11‐mercaptoundecanoic acid; NHS,

N‐hydroxysuccinimide; PBS, phosphate buffered saline; RU, response unit; scFv, single‐chain fragment variable; SPR, surface plasmon resonance.