US 11,692,186 B2
Snake venom thrombin-like enzyme marker peptide of Agkistrodon halys pallas and its application in the species identification of hemocoagulase for injection
Ruiqing Xian, Shandong (CN); Feng Shi, Shandong (CN); Liping Gong, Shandong (CN); Lianli Chi, Shandong (CN); Qunye Zhang, Shandong (CN); Weijian Wang, Shandong (CN); Hongming Du, Liaoning (CN); Congcong Wang, Shandong (CN); Baojian Hang, Shandong (CN); and Fengshan Wang, Shandong (CN)
Assigned to Shandong University, Shandong (CN); Shandong Institute for Food and Drug Control, Shandong (CN); and Avanc Pharmaceutical Co., Ltd, Liaoning (CN)
Filed by Shandong University, Shandong (CN); Shandong Institute for Food and Drug Control, Shandong (CN); and Avanc Pharmaceutical Co., Ltd, Liaoning (CN)
Filed on Jul. 30, 2021, as Appl. No. 17/389,382.
Claims priority of application No. 202010754240.6 (CN), filed on Jul. 30, 2020.
Prior Publication US 2022/0034853 A1, Feb. 3, 2022
Int. Cl. C07K 5/00 (2006.01); G01N 30/72 (2006.01); C12N 9/74 (2006.01); C12Q 1/37 (2006.01); G01N 30/06 (2006.01)
CPC C12N 9/6429 (2013.01) [C12Q 1/37 (2013.01); G01N 30/06 (2013.01); G01N 30/7206 (2013.01)] 3 Claims
 
1. An method of using a snake venom thrombin marker peptide of Agkistrodon Halys Pallas in identifying species of snake venom thrombin for injection, wherein the snake venom thrombin marker peptide of Agkistrodon Halys Pallas comprises the amino acid sequence of SEQ ID NO. 1 and the method comprises the following steps of: A: dissolving the snake venom thrombin for injection and the snake venom thrombin marker peptide of Azkistrodon Halys Pallas into a test solution and a reference solution respectively; B: conducting an alkylation reduction treatment on the test solution and the reference solution with dithiothreitol and iodoacetamide; C: after the alkylation reduction treatment is finished, adding enzyme for a hydrolysis; D: after the hydrolysis is finished, conducting centrifugation at a high speed, and injecting a supernatant into a liquid chromatography-mass spectrometer for analysis; and E: when detecting an ion flow chromatography extracted from ion pairs, if the test solution has a chromatographic peak with a retention time in consistence with the reference solution, indicating that a to-be-detected sample contains the amino acid sequence of SEQ ID NO. 1, and proving that the to-be-detected sample is originated from Agkistrodon Halys Pallas, or otherwise the to-be-detected sample is not originated from Agkistrodon Halys Pallas.