36 results about "Trypsinogen" patented technology

The invention discloses a method for extracting trypsin from animal pancreas. According to the technical scheme of the invention, the zymogen in pancreas cells is extracted by using a sulfuric acid solution; subsequently according to the principle of isoelectric precipitation, the pH is adjusted, and the trypsinogen is deposited and precipitated in a classified salting-in and salting out mode by using ammonium sulfate. By utilizing the method, 6g trypsin can be extracted from 1kg of pancreas.

The present invention relates to a method for the manufacture and purification of recombinant trypsinogen and trypsin in E. coli and yeast, using high yield expression vectors with and without secretion leader sequences. The invention further relates to an improved method and apparatus for carrying out protein refolding specifically useful for processing trypsinogen that has accumulated intracellularly in the form of inclusion bodies.

The invention relates to production and application of high stability recombinant trypsin, and discloses a method for producing the recombinant trypsin, which comprises the following steps of: (1) performing recombinant expression on a fusion protein, wherein the fusion protein sequentially comprises 40-200 amino acid sequences and trypsinogen sequences from an N terminal to a C terminal, and is solubly expressed; and (2) cutting off the 40-200 amino acid sequences and a leader sequence at the N terminal of the fusion protein to obtain the recombinant trypsin. The method realizes the soluble expression of the trypsin, and the high activity and high stability trypsin can be obtained.

The invention discloses a trypsinogen-2 content detection kit and a preparation method thereof. The detection kit consists of reagent I and reagent II which are independent of each other, wherein the reagent I comprises the following components: a biological buffer, a preservative, inorganic salt and water; the reagent II comprises the following components: latex granules coated by trypsinogen-2 antibody, the biological buffer, the preservative and water. The trypsinogen-2 content detection kit adopts the latex granules with the particle size of 100-300nm to prepare the latex granules coated by the trypsinogen-2 antibody, thus effectively guaranteeing the sensitivity of the kit, the detection linearity range, the measurement value accuracy and the bottle opening stability; furthermore, the latex granules coated by the antibody are prepared by a chemical cross-linking method, so that the combination stability between the antibody and the latex granules can be guaranteed, and the stability of the detection kit is improved. The detection kit has the advantages of being good in stability, high in sensitivity, high in specificity, simple and convenient to operate, etc.

The invention relates to a preparation method of recombinant trypsin, and relates to the field of genetic engineering. The invention provides an expression vector and an engineering bacterium comprising trypsinogen. According to the method provided by the invention, plasmids with recombinant trypsinogen gene sequence are introduced into Escherichia coli cells, such that recombinant engineering bacteria are constructed; through fermentation, the expression of recombinant trypsinogen is induced; and through renaturation, enterokinase digestion transformation, and separation purification, trypsin with activity is obtained. The method provided by the invention is characterized in that the recombinant trypsin preparation process is simple, and the cost is low.

The invention relates to a method for expressing a bovine-derived trypsinogen gene in Pichia pastoris by gene recombination. The recombinant bovine trypsinogen is expressed in Pichia pastoris by the novel promoter and secreted into the culture medium; the zymogen is self-activated, or treated by enterokinase or trypsinase or the like to prepare the active bovine trypsinase; and the prepared active bovine trypsinase can be used for producing the recombinant human insulin and human insulin analogs. The method can avoid the defects in the constitutive promoter glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter and the induction-type promoter alcohol oxidase (AOX1) promoter.

The invention provides a production technology of bovine trypsin. By means of a gene recombination technology, bovine trypsinogen or inclusion body protein of bovine trypsinogen fusion protein are expressed out in escherichia coli, and then acidizing is carried out to denature and renature the inclusion body protein into folded bovine trypsinogen or bovine trypsinogen fusion protein. By the adoption of a prokaryotic expression system, the expression quantity is high, denaturation, renaturation and follow-up steps are simple and easy to implement, and the technological cost is low. The prepared bovine trypsin with activity can be used for production of recombinant human insulin and insulin analogues.

The invention provides a Vero cell line for stably secreting and expressing bovine trypsinogen (S.pro-try) and application thereof to influenza virus culture and influenza virus vaccine production. The Vero cell line is named as VTY55, has the preservation number of CGMCC No.12680, and belongs to the technical field of biology. The invention also provides a method for building the Vero-S.pro-try cell line. The Vero-S.pro-try monoclonal cell line VTY55 provided by the invention has the advantages that on one hand, the cell foundation is laid for the large-scale influenza vaccine production in future, and the crucial problems of vaccine productivity expansion and quality improvement can be solved; on the other hand, the cell line can also be used as a study tool in other aspects such as laboratory effective ingredient separation culture, influenza virus monitoring, anti-influenza-virus medicine screening, neutralizing antibody determination and the like; wide application prospects are realized.

The present invention relates to transgenic animal models of chronic pancreatitis. The present invention also provides methods for generating animal models by introducing mutated trypsinogen genes into the germline of animals and screening methods for identifying biologically active compound.

The invention discloses a method for extracting chymotrypsinogen and trypsinogen through fractional precipitation. The principle is that high-concentration saline ions can compete with protein for water molecules in a protein solution so as to damage a hydrated sheath on the surface of the protein and reduce the solubility, and accordingly, the protein can be precipitated from the solution. Ammonium sulfate is most widely used due to large solubility, small temperature coefficient and low probability of protein denaturation. Therefore, the invention adopts an ammonium sulfate fractional precipitation method to research and improve the technology for extracting the chymotrypsinogen and the trypsinogen, the yield of the prepared chymotrypsinogen is improved to 0.5%-0.6% especially by adopting affinity chromatography, and the trypsinogen as a by-product is obtained simultaneously.

The present invention relates to novel Prolipase-Bovine trypsinogen (PLBTR) fusion proteins, the genes encoding them, and the production and uses thereof. More specifically, the present invention relates to methods of producing in optimal quantities PLBTR fusion proteins which comprise a heterologous polypeptide which is normally susceptible to autocatalytic activity. More particularly, the present invention relates to fusion proteins which comprise an heterologous polypeptide, such as a serine protease, fused to a lipase signal sequence, which can be expressed by recombinant host cells in desired amounts. The present invention further relates to polynucleotides encoding such fusion proteins, to expression vectors for expression of such fusion proteins, to host cells transformed with such polynucleotides/vectors, and to methods of generating such fusion proteins.

The invention discloses the application of mogroside IIE in preparing trypsin inhibitors, and belongs to the technical field of medicines. After a long period of extensive trials, the inventor of thepresent application finds that the mogroside IIE can inhibit the activity of trypsin at the acinar cell background, and the inhibition has dependence on time and concentration. In addition, the mogroside IE can inhibit the activation of trypsinogen by an acute pancreatitis inducer-cerulein. At the same time, that the mogroside IE inhibits the activation of the trypsinogen is related to cathepsin B, so that the mogroside IE can be used to prepare trypsin inhibitors.

Pharmaceutical composition containing a mixture of proenzymes and enzymes, containing proenzymes trypsinogen and chymotrypsinogen and enzymes ct-amylase and lipase as active substances, and one or more pharmaceutically acceptable excipients, for simultaneous, separate and subsequent administration of the composition in parenteral or transmucosal way, the composition has anti-proliferative and anti-metastatic effects to cancer tumours and is intended for therapeutic, prophylactic and anti-metastatic use in mammals.

The invention discloses a porcine trypsinogen mutant and an expression of the porcine trypsinogen mutant in pichia pastoris. The porcine trypsinogen mutant is cut by enterokinase to obtain a novel porcine trypsinogen mutant. According to the invention, PGAP is taken as the promoter, methanol induction is avoided, harm of methanol and pollution to the environment are reduced, and the method is safe and convenient. In addition, compared with wild type porcine trypsin, the porcine trypsin mutant has higher stability and activity.

The invention generally provides compositions, kits, pharmaceuticals, and methods that promote and enhance wound healing. Such compositions comprise isolated trypsinogen or trypsin polypeptides or isolated polypeptides obtainable from the excretions/secretions (ES) of Lucilia sericata.

The present invention relates to compositions, methods, uses and kits for treating cancer. The present invention relates to methods for minimising the progression of cancer in a subject, the method comprising administering to the subject therapeutically effective amounts of chymotrypsinogen and trypsinogen, thereby minimising the progression of cancer in the subject. In particular, the methods provide a means for treating cancer by reducing the number of cancer stem cells in the subject.

The invention provides a production process for expressing soluble recombinant trypsin by using yeast, so that trypsin can be highly expressed in the supernatant, and the recombinant trypsinogen can be purified from the supernatant of the fermentation broth by ion exchange chromatography , the highly active recombinant trypsin obtained by autocatalysis is far superior to the Escherichia coli expression process in terms of cost, yield, activity, purity and other technical indicators, so it is of great industrial production value.