40 results about "Limulus" patented technology

Endotoxin, also known as lipopolysaccharides (LPS), is the major mediator of septic shock due to Gram-negative bacterial infection. Chemically synthesized S3 peptide, derived from Sushi3 domain of Factor C, which is the endotoxin-sensitive serine protease of the limulus coagulation cascade, binds and neutralizes LPS activity. Fluorescent tagged-S3 is shown to detect LPS-containing bacteria. For large-scale production of S3 and to mimic other pathogen-recognizing molecules, tandem multimers of the S3 gene were constructed and expressed in E. coli. Tetramer of S3 for example is shown to display an enhanced inhibitory effect on LPS-induced activities. An affinity matrix based on tetramer of S3 is also shown to be particularly efficient at removing LPS.

Endotoxin, also known as lipopolysaccharides (LPS), is the major mediator of septic shock due to Gram-negative bacterial infection. Chemically synthesized S3 peptide, derived from Sushi3 domain of Factor C, which is the endotoxin-sensitive serine protease of the limulus coagulation cascade, binds and neutralizes LPS activity. Fluorescent tagged-S3 is shown to detect LPS-containing bacteria. For large-scale production of S3 and to mimic other pathogen-recognizing molecules, tandem multimers of the S3 gene were constructed and expressed in E. coli. Tetramer of S3 for example is shown to display an enhanced inhibitory effect on LPS-induced activities. An affinity matrix based on tetramer of S3 is also shown to be particularly efficient at removing LPS.

The invention provides a specific limulus reagent and a preparation method thereof. The preparation method comprises steps of blood collection, cell separation, cell lysis, cell wall separation, addition of an activator and a G factor inhibitor, so as to prepare a semi-finished product. The invention further provides a detection kit for blood bacterial endotoxin, wherein the kit contains the specific limulus reagent, and further comprises a humoral treatment agent I, a humoral treatment agent II, bacterial endotoxin controls and inspection water. The specific limulus reagent and the detection kit provided by the invention can qualitatively identify a gel reaction of the limulus reagent with the bacteria endotoxin and glucan type substances, thereby improving reliability of the detection results of the blood bacterial endotoxin, and realizing quantitative detection for the blood bacterial endotoxin. By using the specific limulus reagent and the detection kit, the detection of the blood bacterial endotoxin is characterized by high sensitivity, reliable detection result and high detection efficiency.

The invention relates to a method for using luminous intensity to test the bacterial endotoxin content of glucosum anhydricum, belonging to the analytical chemistry technical field. The technical scheme is that: using luminous intensity to test the reaction time from the reaction of limulus polyphemus agent and the endotoxin to the turbidity of the mixture reaching a preset absorbency; finding the endotoxin content of the object glucosum anhydricum sample. According to the scheme, the method dilutes an endotoxin operation standard sample of known content into operation standard sample series of different contents, tests the times of different endotoxin content standards reaching the preset absorbency, and draws a standard curve; and tests the time of the object glucosum anhydricum sample reaching the preset value, to find the content of the endotoxin of the glucosum anhydricum sample.

The invention relates to limulus egg and maca health-care tablets which comprise the following components according to weight percentage: 4.5%-54% of limulus egg powder, 24%-85.5% of maca powder and the balance of auxiliary materials, wherein the size of particles of the limulus egg powder, the maca powder and the auxiliary materials is 40-100 meshes; and the auxiliary materials comprise microcrystalline cellulose, lactose, sodium carboxymethylcellulose, magnesium stearate and starch. The invention further relates to a preparation method of the limulus egg and maca health-care tablets. The method comprises the following steps of: (1) processing limulus eggs; (2) processing maca; (3) mixing the two processed materials; (4) granulating the mixture; and (5) tabletting the mixture. The limulus egg and maca health-care tablets disclosed by the invention express advantages of eggs of an animal, Chinese limulus, which is taken as both medicine and food, and a plant, maca, which is taken as both medicine and food; the limulus eggs are used for making up the disadvantage that the content of amino acid in the maca is too low, and the maca is used for making up the disadvantages of lack of taurine and low content of a mineral substance zinc in the limulus eggs, so that the aim of making good for deficiency is realized; meanwhile, branched chain amino acids in the limulus eggs and polysaccharide in the maca in promote each other in the aspect of anti-oxidization; and abundant amino acids and unsaturated fatty acids in the limulus eggs are matched with various vitamins, microelements and natural bases of the maca to better improve the immunity, the sexual function and the fertility of people and adjust the internal secretion of people.

The invention discloses a recombinant limulus three-factor reagent which comprises the following components: endotoxin-free water, a heat-resource-free Tris buffer liquid, Tween-20, a fluorescent substrate, a recombinant limulus factor C, a recombinant limulus factor B and a recombinant limulus proclotting enzyme factor. The invention provides the recombinant limulus three-factor reagent and a method for detecting endotoxin with the same, three recombinant limulus factors (the proclotting enzyme factor, the recombinant limulus factor B and the recombinant limulus factor C) can be expressed by using an insect baculovirus expression system, the reagent can be used for detecting bacterial endotoxin, and by adopting the method, bypass interference of a factor G can be eliminated, so that the possibility of false positive caused by glucan is effectively avoided; moreover, the defects that the number of limulus is limited and different batches of limulus reagents are not stable are avoided, and thus the reagent can be used as a novel endotoxin detection reagent.

When assessing the start time of the limulus reaction between biogenous biologically active substances and LAL and using the reaction start time to determine the concentration of the biogenous biologically active substances, in order to exclude the influence of progressive changes which occur regardless of the conditions of the limulus reaction, the strength of transmitted light or scattered light in the liquid mixture of the measurement sample and LAL is detected, the variation (delta) in the transmittance or number of gel particles is acquired at set intervals, and the time when the variation (delta) crosses a threshold value is taken as the reaction start time. Furthermore, the time intervals when acquiring the abovementioned delta are not uniform, and either change over time from the start of measurement as defined by a time function, or multiple sequences with differing time intervals are prepared in advance.

The invention relates to an improved body fluid color-developing fungi 1,3-beta-D-glucan detection kit, which comprises a reaction main-agent, a main agent combination solution, a sample treatment fluid, sterile water, a standard substance and a quality control material. With Tachypleus tridentatus Leach or Limulus polyphemus Linnaeus hemocyte lysate as a main raw material, the reaction main-agent contains G factor, B factor, C factor, clottable protein and a polypeptide chromogenic substrate. By a new preparation process, the reaction main-agent has characteristics of high stability and small intra-assay and inter-assay difference. The main agent combination solution adopts a new formula. After the main agent combination solution is mixed with the reaction main-agent, Tachypleus Amebocyte Lysate endotoxin reaction branch can be effectively shielded. According to the kit, rate-method enzyme kinetics detection is carried out by ELIASA. The detection speed is fast, and anti-interference performance is strong. In addition, the preparation technology is simple, and the product has stronger stability and higher specificity and sensitivity.

It is a blood pre-process method used in horsefoot test, which comprises the following steps: to make the blood pre-process liquid processed by bent X-100 primary liquid, potassium hydroxide, bicine, Ethyleneimineploymer, calcii chloridum and polybrene; to mix it with the serum or blood plasma in the water of 37 degree for eight to fifteen minutes. This method can effectively removes the disturbance materials in the blood sample and reduces the disturbance in the action of the horsefoot and the inner toxin.

The invention relates to a limulus plasma protein hydrolysis method. The limulus plasma protein hydrolysis method comprises the following steps of (1) performing constant-temperature water bath on limulus plasma for 2-4h, sucking and filtering limulus plasma, and washing by water for multiple times for standby use; S2, performing proteolysis on limulus blood obtained in step 1, centrifuging limulus plasma hydrolysate, separating to obtain a supernatant, freezing, and drying, so as to obtain limulus plasma protein hydrolysis peptide powder, wherein the water bath temperature in step S1 is 70-100 DEG C, and the pH (potential of hydrogen) value of a hydrolysis system in step S2 is 7-9. The limulus plasma protein hydrolysis method has the advantages that by utilizing protease to perform enzymatic hydrolysis at most suitable temperature and pH condition, the reaction is mild, the degree of hydrolysis can be controlled, and the nutritional value of amino acid is remained; the experiment equipment is simple, the operation is safe, the pollution by solvent is avoided, and the investment is less.

The invention relates to a preparation method of a protein active peptide of limulus blood hemocytes, wherein the preparation method particularly comprises the following steps: placing the limulus blood hemocytes at low temperature of -18 DEG C, freezing for 12 h, thawing at the temperature of 4 DEG C, repeating three times, and carrying out magnetic stirring at the temperature of 4 DEG C for 10 min until the hemocytes are ruptured; centrifuging at low temperature of 4 DEG C with the rate of 4000 rpm, to obtain a solid precipitate, and repeatedly flushing the solid precipitate with distilled water to obtain a limulus hemocyte membrane tissue; carrying out enzymolysis of the limulus hemocyte membrane tissue with pancreatin powder having the enzyme activity of 4000 u / g, wherein the enzymolysis conditions comprise that the pH is 7.0, the temperature is 60 DEG C-80 DEG C, the enzyme concentration is 2.2 mg / mL, the substrate concentration is 4.0 mg / mL and enzymatic hydrolysis is performed for 5 h; and carrying out centrifugation separation of the enzymatic hydrolysate at low temperature, to obtain a supernatant, and freeze-drying the supernatant, to obtain the protein active peptide ofthe limulus blood hemocytes. According to the method, a residual waste hemocyte membrane tissue for preparing a limulus reagent is used as a raw material, the utilization rate of the raw material is improved, the application scope of limulus blood is enlarged, scavenging of hydroxyl free radicals and acetylcholinesterase is effectively inhibited, and the protein active peptide can be used for treatment and health care of patients with Alzheimer's disease. The preparation method belongs to the technical field of extraction of natural active substances.

The present invention relates to use of a new cell penetrating peptides (CPP) and in particular to the region 32-51 of protein Limulus antilipopolisacárido (LALF) and its analogous. This invention refers to compositions containing these peptides associated to biomolecules with therapeutics properties. This invention consist of compositions comprise the covalent fusion of biomolecules, between this human papillomavirus antigens (HPV) to these CPP for induce a potent immune cellular responses against HPV and HPV protein antigen-exhibiting cells including HPV-associated tumors. The referred compositions are applicable in the pharmaceutical industry as vaccine for therapeutic use in human.

It has been found that a limulus-positive glycolipid is present in xanthan gum derived from Xanthomonas, which has been commercially available and eaten for many years, and this was purified, and it has been found that this limulus-positive glycolipid has an immunopotentiation effect. A method for safely and inexpensively producing the limulus-positive glycolipid containing an immunopotentiator at high concentrations is provided. The method for producing the limulus-positive glycolipid of the present invention comprises extracting the limulus-positive glycolipid from xanthan gum. A limulus-positive glycolipid composition containing the limulus-positive glycolipid can be used for various applications such as pharmaceuticals, pharmaceuticals for animals, quasi drugs, cosmetics, foods, functional foods, feedstuff and bath agents.

A method of eliminating the reactivity of lipoarabinomannan contained in a sample to a Limulus reagent including at least the step of allowing the sample containing lipoarabinomannan to coexist together with a metal salt or a buffer; and a method of assaying an endotoxin and a method of detecting an endotoxin-associated disease by using the above-described method.

The invention relates to a method used for extracting an antibacterial composition from limulus reagent production waste materials. The method comprises following steps: 1, raw material treatment is carried out, wherein a lower layer precipitate waste material obtained in centrifugation of an emulsified product in limulus reagent production is collected, and is taken as a raw material I, and discarded blood plasma in limulus reagent production is collected, and is taken as a raw material II; 2, extraction of a tachyplesin crude extracted solution is carried out, wherein acidolysis is carried out, pH is adjusted so as to remove a precipitate product, and thermal denaturation is carried out; 3, preparation of a hemocyanin crude extracted solution from blood plasma is carried out; 4, preparation of a SOD crude extracted solution is carried out; 5, 60 to 80 parts of the tachyplesin crude extracted solution,10 to 20 parts of the hemocyanin crude extracted solution, and 10 to 20 parts of the SOD crude extracted solution are mixed, pH value is adjusted to 4 to 5, and filtering is carried out so as to obtain the antibacterial composition. According to the method, the crude extracted solutions are prepared rapidly and conveniently at low cost; the antibacterial composition is capable of inhibiting growth of a plurality of kinds of bacteria and fungus; compared with conventional chemical antibacterial products, the antibacterial composition possesses following advantages: the antibacterial composition is safe, mild, and effective, no toxic or side effect is caused, and generation of drug resistance and drug allergy caused by conventional antibacterial products can be avoided.

The invention discloses application of compound lipopolysaccharide of lipid and polysaccharide as a mammal high altitude cerebral edema biomarker. According to the invention, a limulus reagent is utilized for detecting content of lipopolysaccharide in blood plasma of a mammal after entering a high altitude low-oxygen environment, and when the content of the lipopolysaccharide in the blood plasma is more than 0.5EU / ml, content of water in brain tissues of the mammal is obviously increased. The content of the lipopolysaccharide in the blood plasma can be taken as the biomarker that high altitude cerebral edema of the mammal occurs, and an experimental basis is provided for early warning and diagnosing that the high altitude cerebral edema of the mammal occurs.

The invention relates to limulus egg and maca health-care tablets which comprise the following components according to weight percentage: 4.5%-54% of limulus egg powder, 24%-85.5% of maca powder and the balance of auxiliary materials, wherein the size of particles of the limulus egg powder, the maca powder and the auxiliary materials is 40-100 meshes; and the auxiliary materials comprise microcrystalline cellulose, lactose, sodium carboxymethylcellulose, magnesium stearate and starch. The invention further relates to a preparation method of the limulus egg and maca health-care tablets. The method comprises the following steps of: (1) processing limulus eggs; (2) processing maca; (3) mixing the two processed materials; (4) granulating the mixture; and (5) tabletting the mixture. The limulus egg and maca health-care tablets disclosed by the invention express advantages of eggs of an animal, Chinese limulus, which is taken as both medicine and food, and a plant, maca, which is taken as both medicine and food; the limulus eggs are used for making up the disadvantage that the content of amino acid in the maca is too low, and the maca is used for making up the disadvantages of lack of taurine and low content of a mineral substance zinc in the limulus eggs, so that the aim of making good for deficiency is realized; meanwhile, branched chain amino acids in the limulus eggs and polysaccharide in the maca in promote each other in the aspect of anti-oxidization; and abundant amino acids and unsaturated fatty acids in the limulus eggs are matched with various vitamins, microelements and natural bases of the maca to better improve the immunity, the sexual function and the fertility of people and adjust the internal secretion of people.

The invention provides bacterial endotoxin detecting test paper. The test paper comprises a cellulose membrane and a conjugate pad, a bacterial endotoxin antibody is attached onto the cellulose membrane, limulus C factor polypeptide by gene engineering recombination wraps the conjugate pad, and the bacterial endotoxin antibody, limulus C factor polypeptide by gene engineering recombination and bacterial endotoxin make specific combination to form a sandwich structure. The disclosed bacterial endotoxin detecting test paper and kit are independent from natural animal serum and not limited by rawmaterials, the cost can be reduced, the detection process of the test paper and kit is highly automatic, and the test paper and kit are suitable for large-scale production.

The present invention relates to a conjugate of tetrodotoxin (TTX), it contains tetrodotoxin, conjunction agent and carrier covalently-coupled with tetrodotoxin by means of conjunction agent, and thedescribed carrier is selected from Chinese limulus blood blue protein or tetanus toxin or fragment with correspondent biological function. Said invention also relates to the method for preparing saidconjugate, and also relates to the medicine composition containing said conjugate and its application for preventing tetrodotoxism.

The invention discloses a preparation method of a phenoloxidase active protein. The preparation method comprises the following steps: salting out proteins from a raw material waste blood plasma residual on Tachypleus Amebocyte Lysate by a saturated ammonium sulfate solution, performing refrigerating centrifugation to obtain limulus hemocyanin, performing dialysis desalting on a PBS solution of thehemocyanin through an MW 3500 Da dialysis bag, and carrying out Sephacryl S-100 allyl dextran gel column purification and concentrating treatment to obtain high-efficiency phenoloxidase active hemocyanin, wherein the initial reaction rate Vi of the high-efficiency phenoloxidase active hemocyanin is 74.52 nmol.(min.mg). The product obtained in the invention has a high purity and a high activity, and can be used for preparing a phenolic pollutant micro-detector with the advantages of high efficiency, simplicity and low cost; and the preparation method is simple, and simultaneously achieves desalting, separating, purifying and concentrating effects by cooperating the MW 3500 Da dialysis bag with the Sephacryl S-100 allyl dextran gel column.

The invention discloses broiler feed capable of reducing emission of hydrogen sulfide. According to the broiler feed, corn is selected as an energy raw material, and a part of soybean meal is replaced with cottonseed meal and peanut kernel meal in a proper proportion. The use amount of sulfur-containing amino acid methionine is reduced, and the synthesis action of an animal body is enhanced by adding betaine and increasing the use amount of reinforced choline chloride, folic acid and the vitamin B12, so that growth needs are met. By adding an enzyme preparation, prebiotics and antibacterial peptide (limulus), the immunity of the intestinal tract and the health level of the intestinal tract are improved, and the absorption and utilization rate of the intestinal tract to nutrients including sulfur is promoted. Therefore, the developed feed can promote the growth of broilers, improve the sulfur utilization rate of the feed and reduce emission of hydrogen sulfide in the breeding process.

The invention relates to the detection for bacterial endotoxin, concretely a limuloid reagent sensitivity determining method, its character: 1) adopt standard endotoxins at 3-6 concentrations to test standard limuloid reagents at different sensitivities on the endotoxin determiner, make linear regression on the logarithms of reacting time and endotoxin concentration, to obtain the corresponding standard curves at different sensitivities as the justification basis; 2) make the limuloid reagent to be measured react with standard endotoxins at 1-6 concentrations, on the corresponding condition of reacting time or solution absorbency, use the standard curves to calculate endotoxin concentration and recovery, according to the sensitivity corresponding to the standard curve at the recovery 75-120%, determine it as the sensitivity of the limuloid reagent to be measured.

A method of eliminating the reactivity of lipoarabinomannan contained in a sample to a Limulus reagent including at least the step of allowing the sample containing lipoarabinomannan to coexist together with a metal salt or a buffer; and a method of assaying an endotoxin and a method of detecting an endotoxin-associated disease by using the above-described method.

The invention relates to the detection for bacterial endotoxin, concretely a limuloid reagent sensitivity determining method: 1) adopt standard endotoxin at the same concentration to repetitively test standard limuloid reagents at different sensitivities on the endotoxin determiner, to obtain a average value and standard difference of the corresponding characteristic reacting time; 2) make the limuloid reagent to be measured react with the above standard endotoxin to obtain a corresponding characteristic reacting time, then compare the characteristic reacting time whit the above average value and standard difference to judge and obtain the limuloid reagent sensitivity.

Provided are all full-length recombinant proteins involved in the clotting mechanism of Limulus polyphemus, cDNAs encoding the same, and applications thereof. A recombinant protein containing the amino acid sequence represented by SEQ ID NO: 2 or 4, a recombinant protein containing the amino acid sequence represented by SEQ ID NO: 6, 8, 10, or 12, a recombinant protein containing the amino acid sequence represented by SEQ ID NO: 14, 16, 18, 20, or 22, variants thereof, cDNAs encoding the same, and utilization thereof.

The invention relates to the detection for bacterial endotoxin, concretely a limuloid reagent sensitivity determining method, its character: 1) adopt standard endotoxins at 3-6 concentrations to test standard limuloid reagents at different sensitivities on the endotoxin determiner, make linear regression on the logarithms of reacting time and endotoxin concentration, to obtain the corresponding standard curves at different sensitivities as the justification basis; 2) make the limuloid reagent to be measured react with standard endotoxins at 1-6 concentrations, on the corresponding condition of reacting time or solution absorbency, use the standard curves to calculate endotoxin concentration and recovery, according to the sensitivity corresponding to the standard curve at the recovery 75-120%, determine it as the sensitivity of the limuloid reagent to be measured.