32 results about "Agkistrodon blomhoffii" patented technology

The invention provides a novel snake venom polypeptide, which is extracted from the coarse venom of pit vipers and has molecular weight of 7554Da. The sequence of 1-15 amino acids at the N terminal is shown in formula (I), wherein E is glutamic acid; A is lactamic acid; D is aspartic acid; N is asparagines; C is cystine; and N represents an amino terminal. The snake venom polypeptide has high anti-tumor activity, and is applicable in preparing anti-tumor pharmaceuticals. The formula (I) is N-EAEEDEDDDAAAANC.

The invention provides a kit for detecting circulating antigens of Agkistrodon acutinae using specific polyantibody, comprising rabbit anti-Aag-IgG, enzyme-labeled antibody (HRP-Aag-IgG), nitrocellulose film, 1 % bovine serum albumin in PBST solution, PBST washing solution, substrate chromogenic solution, positive and negative control sample composition and polyethylene reaction plate. The invention also provides a preparation method of a kit for detecting circulating antigens of Agkistrodon acutina by using specific polyantibody. The invention adopts the polyclonal antibody to measure the circulating antigen, which is beneficial to making early diagnosis and helping to judge the prognosis. When the parasites in the body die, the circulating antigens disappear quickly, which can be used for efficacy assessment. The kit of the invention has high sensitivity and strong specificity, and can realize rapid diagnosis, drug screening and curative effect assessment of Agkistrodon acutinae ligamentiworm disease.

The invention provides agkistrodon acutus hemocoagulase-B which is high-activity hemocoagulase separated from agkistrodon acutus venom. The hemocoagulase is single-stranded glycoprotein comprising 236 amino acids, and has an amino acid sequence shown as SEQ ID No. (sequence identification number) 1. The strand comprises 6 pairs of disulfide bonds; molecular weight of SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) is approximately 35kD; the molecular weight of the hemocoagulase after being deglycosylated is 26116.7Da; and an isoelectric point (pI) is 6.0. A hemocoagulase molecule is modified by heterozygotic polysaccharides at an Asn<77, 100, 229> (asparagine) site. The hemocoagulase is serine proteinase. The invention also provides a separation and purification method of the hemocoagulase, which comprises the steps that undissolved substances are removed through pretreating; then two times of anion-exchange column chromatography and one time of Sephdex-G75 molecular sieve chromatography are conducted; an active elution peak is acquired; dialysis and lyophilization are conducted; and the high-purity agkistrodon acutus venom hemocoagulase is obtained. The specific activity of the agkistrodon acutus venom hemocoagulase is not less than 2000U/mg; the analysis purity of HPLC (high performance liquid chromatography) can reach above 95%; and the yield is 0.25-0.30% calculated according to the weight of an agkistrodon acutus venom raw material.

The invention discloses a method for identifying agkistrodon halys venom by applying mass spectrometry. The method comprises the following steps of: (1) dissolving agkistrodon halys venom to prepare aprotein solution; (2) carrying out SDS-PAGE analysis; and (3) carrying out LC-MS/MS mass spectrometry to carry out agkistrodon halys venom component identification. In addition, the invention furtherdiscloses application of the identification method in the preparation of products for identifying the agkistrodon halys venom and application in the preparation of agkistrodon halys venom resistant serum for treating agkistrodon halys venom poisoning. According to the invention, the agkistrodon halys venom can be identified in a laboratory; agkistrodon halys venom is more accurately subjected toquality control, so that specific drug anti-agkistrodon halys poison serum for treating agkistrodon halys venom poisoning is produced; and the method is very important for ensuring the safety, effectiveness and quality controllability of anti-agkistrodon halys venom serum products and is of great significance for reference of the diagnosis and legal medical expert identification of the types of venom caused by snake venom poisoning. The method has the advantages of high detection sensitivity, high correct detection rate, no cross reaction and simple operation process.

The invention belongs to the field of molecular biology and particularly relates to an anti-inflammatory polypeptide DAvp-1 in snake venom and application of the anti-inflammatory polypeptide DAvp-1. The invention specifically provides an anti-inflammatory polypeptide DAvp-1 with a polypeptide sequence as shown in SEQ ID NO: 2. According to the anti-inflammatory polypeptide DAvp-1 and the application thereof, a brand-new polypeptide DAvp-1 capable of being combined with TNFR1 is successfully screened from a phage library of venom of deinagkistrodon acutus. The polypeptide DAvp-1 has a potential effect of antagonizing TNF-alpha and is an excellent candidate for research and development of polypeptide anti-inflammatory drugs.

The invention relates to a preparation method of a recombinant viper venom protein. A mutant Alfimeprase gene of artificially synthesized Agkistrodon contortrix is utilized, and the gene has the nucleotide sequence shown as SEQ ID No.2. The gene is linked to yeast expression plasmids pPIC9K to construct a high efficiency expression vector pPIC9K-His-Alf, a Pichia yeast strain GS115 is introduced, and the vector is named as FY-Alf. Induced expression is carried out with methanol, the expressed recombinant protein is purified and then subjected to activity verification by a fibrin plate assay, and the result shows that the recombinant protein Alfimeprase expressed by the yeast strain GS115 containing pPIC9K-His-Alf plasmids has high fibrinolytic activity.

The invention provides a method for purifying agkistrodon spearhead hemocoagulase, which comprises the following steps: pretreating agkistrodon spearhead venom, and sequentially carrying out benzamidine agarose gel affinity chromatography, cation exchange chromatography and hydrophobic chromatography to obtain the agkistrodon spearhead hemocoagulase. According to the method provided by the invention, the process steps are further simplified, the purification period of the hemocoagulase is greatly shortened, the process environment exposure risk is reduced, the process stability is high, the product quality consistency is good, and the clinical medication safety is facilitated.

The invention relates to the technical field of biochemical detection, and particularly discloses an agkistrodon blomhoffii ussurensis defibrase HPLC detection method and application thereof. According to the HPLC detection method, a mobile phase A, a mobile phase B and a mobile phase C are adopted for elution; the mobile phase A is a 20-25 mM aqueous solution of sodium dihydrogen phosphate, and the pH value of the aqueous solution of sodium dihydrogen phosphate is 5.8-7.0; the mobile phase B is acetonitrile; and the mobile phase C is an aqueous solution of trifluoroacetic acid. Octadecylsilane chemically bonded silica is used as a filling agent in a reversed-phase column, and gradient elution is carried out. By establishing a purity determination method which is simple, convenient and sensitive and has good specificity, accuracy and durability, the quality of the agkistrodon blomhoffii ussurensis defibrase can be effectively controlled, and the quality of the agkistrodon blomhoffii ussurensis defibrase is stable, controllable, efficient and safe. Meanwhile, the method can also be used for identifying defibrases from different snake venom sources, and lays a foundation for improving quality standards of defibrase raw materials and preparations thereof.

The invention discloses a traditional Chinese medicine for preventing and treating cerebral arterial thrombosis. The traditional Chinese medicine comprises the following raw material components in mass ratio: 1000 of agkistrodon halys powder, 1000 of radix astragali powder, 500 of caulis spatholobi powder, 100 of hirudo powder, 100 of pheretima powder, 100 of rhizoma chuanxiong powder, 100 of radix salvia miltiorrhiza powder, 100 of rhizoma gastrodiae powder and 1 of moschus powder. Agkistrodon halys and radix astragali are monarch drugs and have the effects of tonifying qi and activating blood; rhizoma gastrodiae, hirudo and radix salviae miltiorrhizae are used as ministerial drugs for eliminating phlegm and removing blood stasis; rhizoma chuanxiong, caulis spatholobi, pheretima and moschus are assistant and guide each other to activate blood and promote blood circulation and enter brain orifices; and all the medicines are combined to achieve the effects of promoting blood circulation, removing blood stasis, tonifying deficiency and dredging collaterals, and the traditional Chinese medicine is safe to take and free of side effects, and can be used for preventing and treating cerebral arterial thrombosis.