US 11,874,275 B2
Method for fluorescent immunoassay (FIA) and chemiluminescent immunoassay (CLIA) based on electrokinetic acceleration
Xiaozhu Liu, Guangdong (CN); Hai Xu, Guangdong (CN); Yanmin Li, Guangdong (CN); Jun Li, Guangdong (CN); Yong Hu, Guangdong (CN); Li Tong, Guangdong (CN); Jie Lin, Guangdong (CN); Zhidong Zhang, Guangdong (CN); Lihua Yang, Guangdong (CN); Liang Ma, Guangdong (CN); Zheng Zeng, Guangdong (CN); Linggao Zeng, Guangdong (CN); Li Chen, Guangdong (CN); Shengxi Wu, Guangdong (CN); and Shenghui Qin, Guangdong (CN)
Assigned to FOSHAN MICROWONDERS BIOTECHNOLOGY CO., LTD., Foshan (CN)
Filed by FOSHAN MICROWONDERS BIOTECHNOLOGY CO., LTD., Guangdong (CN)
Filed on Jun. 5, 2023, as Appl. No. 18/205,581.
Application 18/205,581 is a continuation of application No. PCT/CN2022/085472, filed on Apr. 7, 2022.
Claims priority of application No. 202110517207.6 (CN), filed on May 12, 2021.
Prior Publication US 2023/0324380 A1, Oct. 12, 2023
Int. Cl. G01N 31/00 (2006.01); G01N 33/53 (2006.01); G01N 33/557 (2006.01); G01N 33/543 (2006.01)
CPC G01N 33/557 (2013.01) [G01N 33/5438 (2013.01)] 16 Claims
 
1. A method for fluorescent immunoassay (FIA) and chemiluminescent immunoassay (CLIA) based on electrokinetic acceleration, comprising the following steps sequentially:
S1. sample acceleration, comprising: dripping a target molecule on a chip and applying an actuating signal to obtain a chip binding with the target molecule, wherein the chip comprises an electrode sheet, coating molecules is immobilized on the electrode sheet, and the coating molecule is provided for binding specifically to the target molecule;
S2. secondary antibody acceleration, comprising: adding a secondary antibody for binding to the target molecule dropwise on the chip binding with the target molecule, and applying an actuating signal to the chip to obtain a chip binding with the secondary antibody; and
S3. detection, comprising: subjecting the chip binding with the secondary antibody to fluorescence detection or chemiluminescence detection,
wherein during the sample acceleration of S1, a preparation process of the chip comprises a chip coating step for immobilizing the coating molecule on the electrode sheet of the chip, wherein at a specific impedance scanning frequency, relative to an electrode sheet not immobilized with coating molecules, the electrode sheet immobilized with the coating molecule has a capacitance change rate of −50% to 150%, an impedance change rate of −100% to 100%, a phase change rate of −30% to 30%, a resistance component change rate of −40% to 40%, or an inductance component change rate of −200% to 200%;
the chip coating step comprises preparation of coating molecules solution with a boric acid buffer as a solvent; the boric acid buffer is prepared as follows: adding a 0.0125 M to 0.05 M sodium tetraborate solution to a 0.05 M to 0.2 M boric acid solution until a pH is 5 to 8; and a material of the electrode sheet is aluminum.