Biotechnology drugs mainly include polypeptide drugs, protein drugs and nucleic acid drugs. Compared with chemically synthesized drugs, it has high molecular weight, poor stability and low bioavailability after administration. Therefore, how to use modern pharmacy and preparation technology to prepare new preparations of biotechnological drugs to ensure the safety and effectiveness of drug use has become a research hotspot in the field of biotechnological drugs and pharmacy.
Since 1980s, biotechnological drugs have made great progress and become the key projects of China's development, and gradually developed from biological products, biochemical drugs and microbial drugs to biotechnological drugs. China's bioengineering pharmaceutical industry is a new industry, which is far from developed countries. However, the research and development of products has taken a shortcut and has the characteristics of fast start. From 1989, the first bioengineering drug in China, to 2003, there were 2 1 genetically engineered drugs in China, including recombinant human interferon, erythropoietin, interleukin -2, human auxin, staphylokinase, recombinant human tumor necrosis factor, nerve growth factor and human insulin, among which nearly/kloc- Biotechnology drugs have become an important category in the pharmaceutical market, which are used to treat cancer, human immunodeficiency virus diseases, cardiovascular diseases, diabetes, anemia, autoimmune diseases, genetic defects and many genetic diseases. With the rapid development of bioengineering technology, biopharmaceuticals will surely become the fastest growing part of the pharmaceutical industry.
Method for establishing phage polypeptide fluorescent protein labeling system
Project Description: The present invention relates to a screening method of promoter activator of aging inhibition gene, in particular to a screening method of Klotho gene promoter activator, belonging to the technical field of molecular pharmacology. The screening method comprises the following steps: connecting a Klotho gene promoter with a reporter gene, then transfecting the connected gene into mammalian cells to form a stable cell strain, adding an oxidant into the stable cell strain to establish an aging cell model, then adding a traditional Chinese medicine extract to be screened, measuring the expression level of the reporter gene, evaluating the biological activity of the traditional Chinese medicine, and separating the traditional Chinese medicine extract with high biological activity into a single compound to obtain a Klotho gene promoter activator. This is a sensitive screening method, which can be applied to the activation amount of Klotho gene promoter in Qualcomm.
Method for inducing/preparing epitope-specific monoclonal antibody by epitope polypeptide
Project Description: The invention belongs to the technical field of bioengineering. Firstly, an epitope polypeptide with a specific epitope on a natural protein or a gene recombinant protein is artificially synthesized; One or more epitopes often appear on synthetic peptides (i.e., epitope polypeptide; Its epitope polypeptide is coupled with carrier protein or carrier polypeptide; The epitope polypeptide conjugate is matched with an appropriate adjuvant to immunize animals; Hybridoma were prepared by conventional cell fusion technique. The invention can efficiently and directly induce/prepare that specific monoclonal antibody of the predetermine epitope without natural protein or gene recombinant protein.
Application of chitosan oligosaccharide and its derivatives as drug carriers for anti-HIV- 1 polypeptide
Project Description: The purpose of this invention is to provide the application of chitosan oligosaccharide and its derivatives as anti-HIV- 1 polypeptide drug carriers. Chitooligosaccharides are oligoglucosamine made from 2-20 glucosamine obtained by degrading chitosan by bioengineering technology. It has the important functions of enhancing immunity, reducing blood fat, lowering blood sugar, preventing cancer cell metastasis, and inhibiting cell aging. As a carrier of polypeptide drugs, chitosan oligosaccharide can increase the bioavailability of polypeptide drugs with low absorption rate; Controlling the release of polypeptide drugs; Reduce the irritation to gastrointestinal mucosa; Maintain that stability of polypeptide drugs in vivo; Improve the targeting of drugs.
Study on the Construction and Function of Advanced Structures of protein and Polypeptide
Project Description: This project takes the hydrogen bond system formed by protein and polypeptide as the research object, takes model design and theoretical calculation as the exploration means, and takes the construction of advanced structure, functional research and essential exploration as the basic goals. Through three years' practice, we have a new understanding of the reasons why insect antifreeze proteins TmAFPs recognize each other to form dimers, the role of crystalline water molecules in the process of dimerization and protein-ice orientation, and the relationship between the number of β rings in proteins and antifreeze activity. The synergistic effect of superaddition is the internal reason why β -type cyclic peptides can form stable polypeptide nanotubes through self-assembly, and the substituents of D and L-type cyclic peptides are the important determinants of their self-assembly in parallel or antiparallel manner through intermolecular hydrogen bonds. When azobenzene groups are introduced into cyclic polypeptides such as [(-L-phe1-d-men-ala2) 4-], molecules can interact through hydrogen bonds. These works provide basic results for the construction and function of advanced structures of protein and peptides.
A New Class I Genetic Engineering Drug —— Antithrombotic Polypeptide
Project Overview: This project is a national first-class new drug with independent intellectual property rights based on the latest coagulation mechanism. Our research group developed a powerful antithrombotic polypeptide by genetic engineering technology, which has a strong inhibitory effect on venous thrombosis and arterial thrombosis in vivo. The effective dose is lower than 1mg/kg (single subcutaneous injection), which can be used for various thromboembolic diseases such as myocardial infarction.
At present, the construction and screening of genetically engineered bacteria, the stability study of strains, the optimization of protein expression conditions, the purification of proteins, the pharmacodynamics study and the preliminary toxicity study have been completed. Compared with heparin anticoagulants currently used in clinic, it has obvious advantages such as strong efficacy, convenient administration, long duration of efficacy, light adverse reactions, good stability and simple purification process, and has great market potential in the future.
Long-acting blood glucose regulating polypeptide
Project outline: Long-acting blood sugar regulating polypeptide belongs to 1 class genetic engineering new drug, which is used to treat diabetes. Its main features include three aspects: first, long-term, once every two days; Second, no side effects, normal blood sugar adjustment, will not cause hypoglycemia; Third, it does not form drug resistance.
Fully automatic polypeptide synthesizer
Project Overview: The project has developed super-large, super-small and medium-sized polypeptide synthesizer, which meets the needs of large-scale polypeptide production in the factory and micro-research in the laboratory, abandons the design defects of existing products, greatly improves the efficiency of polypeptide drug production and research and development, and fills the gap in China. The only polypeptide synthesizer that meets FDA certification requirements; Fully linear amplification; Fully stir, leaving no dead ends; The recycling of reagents can save about 40% of reagents; Fault alarm and personal safety protection. An operating system that meets the requirements of FDA for GMP production conditions of instruments and equipment; Linear amplification reactor accelerates product development cycle; 0 ~ 180 degrees up and down rotating stepless speed regulation mixing system to eliminate dead angle of reaction; The reagent recovery system can save about 40% reagents; Unique pipeline design eliminates cross contamination of raw materials or reagents; The design of software and hardware fully considers the characteristics of other solid-state reactions except peptides, so that one machine can be used for multiple purposes.
Analysis of Polypeptide Components of Jilin Ginseng
Project Description: The main feature of this project is the combination of organic synthesis technology and microextraction technology, which can be called reactive extraction method for the study of peptides in ginseng, a rare Chinese medicine in Jilin. The preparation, extraction efficiency and selective extraction of polypeptide by extractant were studied. The immobilization of peptide extraction reagent and the method of peptide extraction were studied. The biological activity, quantitative analysis and structural characterization of the extracted polypeptide were carried out. The immobilized peptide extraction reagent is recycled.
Directional bifunctional anti-tumor polypeptide
Project Description: The targeted bifunctional anti-tumor polypeptide with independent intellectual property rights developed in this project is a hybrid polypeptide molecule formed by connecting two polypeptide molecules with different functions through a carrier, which not only has the function of targeting and inhibiting the growth of tumor cells for a long time (only four injections every three days, the tumor inhibition rate of mouse lung cancer is over 80%), but also can inhibit the metastasis of tumor cells (only one intravenous injection is needed to inhibit mouse B 16 melanoma). Low effective dose and extremely low toxicity.
Regulation of bionic immobilization of polypeptide on specific adhesion of transgenic bone marrow stromal cells
Project Description: In this study, GRG DSPC, the best polypeptide to mediate the efficient adhesion of bone marrow mesenchymal stem cells (BMSC), which is the best seed cell in bone tissue engineering, was designed and screened, and then it was immobilized on a porous matrix prepared from poly (lactide-glycolide)-aspartic acid-polyethylene glycol PLGA(ASP-PEG) triblock * * * with nanometer spacing. The combination of peptide and PLGA can be strengthened by * * * polymerization modification, and the aggregation state of integrins on cell membrane surface can be regulated by controlling the density and spatial distribution of polypeptide, thus mediating the specific adhesion of BMSC more effectively, and solving the incompatibility problem of seed cell-material interface to some extent.
At the same time, transforming growth factor β 1(TGF-β 1) gene was transferred into bone marrow mesenchymal stem cells, which were compounded with bionic matrix materials to construct tissue engineering artificial bone with good biological activity. TGF-β 1 gene with multiple biological effects can regulate the directional differentiation of seed cells more continuously and efficiently, and reveal its signal transduction mechanism for the first time. Therefore, it lays a good foundation for developing more ideal new bone repair materials, which has great scientific significance and broad clinical application prospects.
Relationship between primary structure of polypeptide chain and chemical properties of free radicals
Project Description: Among the constituent units of protein molecule, 20 kinds of amino acids belong to functional organic small molecules. These small biomolecules with different structural and functional properties form protein biomacromolecules with diverse structures and functions through the connection of peptide bonds and the change of sequences.
Based on the principles of organic chemistry and biology, this project explores the molecular mechanism of the molecular transformation of reducing protein and the relationship between reducing amino acids and the molecular function of protein through in-depth exploration and systematic integration of various physical and chemical properties of amino acids.
Application of enzymatic hydrolysis of polypeptide amino acids in food seasoning
Project Description: This study uses biotechnology to extract high-quality amino acid solution from waste beer yeast mud, which has no peculiar smell, rich nutrition and good application effect. Extracting amino acid solution from the leftovers of beer production is not only low cost, but also beneficial to environmental protection and cleaner production. For the first time, yeast extract with health care function was strengthened in aquatic algae seasoning. At the same time, pure natural substances are used to prevent the loss of nutrients in the product. Become a real green functional food and healthy food.
Obtaining and Application of Polypeptide for Inhibiting Cell Growth
Project Description: This technology involves a polypeptide consisting of 25 amino acids (methionine-aspartic acid-arginine-aspartic acid-alanine-aspartic acid-tryptophan-arginine-glutamic acid-valine-methionine-proline-tyrosine-serine-threonine-glutamic acid-leucine-phenylalanine-tyrosine.
Study on the Structure and Function of Polypeptide Fragment of Myosin PEVK
Project Description: Based on the amino acid sequence analysis results of myosin PEVK fragment, the molecular conformation analysis of EVPK, KVPE and corresponding phosphorylated tetrapeptide was carried out. The analysis shows that the low-energy conformational skeleton can be divided into two groups: folding and stretching, which shows that the molecule can be elastic in macroscopic view. Phosphorylation has little effect on molecular skeleton, so the elasticity of PEVK polypeptide fragment may not be controlled by phosphorylation.
In this project, molecular simulation is used to simulate the structure and properties of unknown or not yet synthesized protein, and the structure and function of the whole protein are explained by studying the structure and properties of the motif. The results will provide important experimental and theoretical basis for explaining the elastic function of PEVK polypeptide fragment in myosin and muscle from the molecular point of view.
Novel multifunctional anti-inflammatory and anti-tumor polypeptide
Project Description: Many inflammation-related diseases are common diseases that are difficult to cure in clinic. Transcription factor NF-kB is the key switch for the body to induce inflammatory response, and it is considered as the golden target for developing anti-inflammatory drugs. In this project, several highly effective and specific bioactive peptides targeting NF-kB were screened. In vitro and in vivo experiments have proved that these peptides have anti-inflammatory and anti-tumor effects with high efficiency and low toxicity, and have good market prospects and social and economic benefits. These peptides have applied for 4 patents and successfully established a mature and stable polypeptide production process. At present, it is necessary to increase investment, speed up its preclinical research and industrial production, and apply for clinical approval of new drugs. In order to complete the preclinical research, investors need to invest RMB 3 million-5 million separately or jointly; After obtaining the clinical approval, the investor can continue to conduct clinical research on new drugs solely or at the expense of the investor.
Study and Application of Biotransformation Extraction Technology of Yak Bone Marrow Protein Polypeptide Osteogenic Protein
Project Overview: This project takes yak bone, a characteristic resource in Qinghai-Tibet Plateau, as raw material, and uses modern biotechnology and modern food and drug processing technology to systematically study the extraction of yak bone marrow polypeptide bone protein, establish corresponding production process model and process parameters, and study yak bone marrow polypeptide bone protein products.
In this study, yak bone was pretreated by high temperature and high pressure cooking, supplemented by various biological enzyme hydrolysis methods, which not only improved the yield of protein polypeptide collagen, but also retained the nutritional components and improved the product quality. The process conditions are mild, efficient and specific, the quality is safe and reliable, and the required instruments and equipment are simple and easy to operate.
In this study, the superfine powder of tremella fuciformis was used as the auxiliary material of bone collagen, which successfully solved the problems of easy moisture absorption, poor fluidity and insufficient taste of bone collagen products, enhanced the health care function of the products, was innovative and filled the gap in this field.
Research progress of anti-inflammatory peptides
Project Description: Intercellular adhesion molecule-1( 1CAM- 1) mediates the mutual recognition and adhesion between cells, and its over-expression and disorder will cause many diseases. The antagonistic peptide of ICAM- 1 can be used to treat various inflammations in clinic. Screening target functional peptides from display peptide library is a high-tech that has attracted much attention in the field of biotechnology and pharmacy.
After several rounds of affinity panning, a pentapeptide was obtained from this project, which can specifically bind ICAM- 1. Experiments show that it can block the signal transduction pathway of inflammation. This study is expected to provide new and more effective drugs for the treatment of inflammation. Small peptide itself has no immune reaction and toxic side effects, is safe and reliable, and can be used to treat various inflammations.
Production technology of high nutritional peptide milk and peptide milk powder
Project Description: Polypeptide fresh milk and milk powder are made from milk. Through scientific biotechnology treatment, protein macromolecules such as casein in milk (molecular weight is usually hundreds of thousands to hundreds of thousands) are converted into peptides with molecular weight of 1000 to 8000Da. This kind of fresh milk and milk powder can be used to maintain and improve the nutritional status of protein, which is easy to digest and absorb. On the one hand, it can be provided to people with weak gastrointestinal function, such as convalescent patients, elderly people with decreased digestive function, infants with immature digestive system, etc. On the other hand, it can be used as an efficient supplement to protein after excessive fatigue, especially for the rapid recovery of athletes' physical fitness after strenuous exercise. Through scientific processing and transformation, polypeptide fresh milk and milk powder also contain some active peptides with good physiological effects on human body, which is beneficial to human body's absorption of calcium, promotes growth, and has the functions of lowering blood pressure and blood lipid.
The technology of this project is reasonable and mature, which solves the technical problems such as bitterness and browning that are easy to occur in the production process. Dairy factories and milk powder factories above medium scale can take over production, and they can be put into production only by slightly adding some equipment.
Screening and preparation of polypeptide drugs by phage display technology
Project Description: Using phage display technology to screen functional peptides, which can be used as a variety of human growth factors (nerve growth factor family, polypeptide hormones, other cytokines, etc. ) or as a targeting peptide for biological missiles, has become one of the most cutting-edge and hot topics in the international medical field. And has great application prospect. Its research basis is: 1. Phage library has been established, including 6 amino acid random peptide libraries and 15 amino acid random peptide libraries. 2. First-class animal experiment, tissue culture and cell culture conditions. 3. With the research accumulation of relevant national natural science funds and key national research projects in the Ninth Five-Year Plan, it has a high level in research technology and researchers' staffing.
Application scope and market forecast of this project: As the guiding peptide of biological missiles, it is one of the most advanced and hot topics in the medical field in the world today. It has a good application prospect.
Expression and activity of recombinant human lysozyme and bovine lactoferrin N-terminal polypeptide gene
Project outline: The renaturation conditions of recombinant fusion protein pGEX-4T 1/rbLF-N inclusion bodies were explored, and the optimum renaturation conditions were determined as follows: pH 8.5, protein solution concentration 100μg/mL, DTT concentration 24mmol/L, and the renaturation rate of inclusion bodies was the highest. The fusion protein GST-rbLF-N purified by affinity chromatography was identified by SDS-PAGE analysis. The purity of target protein rbLF-N obtained by thrombin digestion was 93.23% by HPLC. The components of GST-rbLF-N digested by thrombin and pepsin were identified and analyzed by LC-MS/MS, and it was proved that GST-RBLF-N contained the active components of natural bLFcin. The recombinant protein rbLF-N was hydrolyzed by pepsin, and the activity of G+ bacteria and G- bacteria was detected, which proved that it had antibacterial activity.
In this paper, the recombinant peptide was concentrated, separated and purified by a membrane with molecular weight of 5kDa. The membrane flux increases with the increase of system temperature. Considering the operating cost and irreversible denaturation of recombinant protein, the suitable operating temperature is 32℃. In this paper, the recombinant expression system of N-terminal polypeptide of bovine lactoferrin was established, and the antibacterial activity of the recombinant N-terminal polypeptide of bovine lactoferrin was determined, which proved that it had natural antibacterial activity.
Human alpha1-thymosin complex and preparation method thereof
Project Description: The complex is composed of human α 1- thymopeptide and monomethoxy polyethylene glycol, which are respectively connected with succinimide ester bond or aldehyde group of monomethoxy polyethylene glycol through ε-NH2 or N-terminal α-NH2 of lysine in human α 1- thymopeptide to form stable amide bond or secondary amine bond. The free amino group on the surface of human α 1- thymopeptide protein reacts with succinimide-activated monomethoxypolyethylene glycol with active group or monomethoxypolyethylene glycol with aldehyde group, and the two are connected to form monomethoxypolyethylene glycol-human α 1- thymopeptide complex, that is, mPEG-Tα 1.
The preparation method of the invention is simple and convenient to operate, and the raw materials are easily available. The compound prepare by that invention has high biological stability and long in-vivo half-life, and is beneficial to improving the bioavailability of human alpha1-thymosin.
Human glucagon-like peptide-1 complex and preparation method thereof
Project Description: In this study, the recombinant human glucagon-like peptide-1(GLP- 1) was efficiently expressed by genetic engineering technology, and the recombinant human glucagon-like peptide-1 was modified by PEG modification technology to prepare PEG-GLP- 1 complex, which has good biological stability, prolonged half-life and antigenicity in vivo. Animal experiments show that the effective hypoglycemic time of this compound is 6 times longer than that of natural GLP- 1. The results are characterized by lowering blood sugar and body weight, without hypoglycemia, which lays a good foundation for the preparation of long-acting immune enhancers.
Research and Development of a New Generation of Broad-spectrum and High-efficiency Antibacterial Peptides
Project Description: This project is a new generation of antibiotics with independent intellectual property rights-antimicrobial peptides. At the end of 2005, this new antibacterial peptide was declared as an international patent of the World Patent Cooperation Organization. At present, there are two new varieties of high-efficiency broad-spectrum antibacterial peptides, and related preclinical research and application work are being carried out.
The project is committed to manufacturing real broad-spectrum antibacterial peptide drugs for injection, and producing and selling them. The product of this project is a brand-new broad-spectrum antibiotic with great market share and prospect.
Clinical study on detection of glomerular filtration function by anti-sulfhydryl protease peptide C
Project Description: This experiment studied the serum cystatin C concentration of children and adults. The concentration of serum cystatin C in children after the age of l is stable and has no correlation with serum creatinine. The concentration of serum cystatin C in adult group is stable, which is positively correlated with serum creatinine and renal function damage. /kloc-There is no difference in serum cystatin C between children over 0 years old and adults, and serum cystatin C has nothing to do with age, height, weight and sex. Serum cystatin C can reflect glomerular filtration rate more accurately and serve the clinic better. Serum cystatin C has a broad clinical application prospect.
Application of chitosan oligosaccharide and its derivatives as drug carriers for anti-HIV- 1 polypeptide
Project Description: The purpose of this invention is to provide the application of chitosan oligosaccharide and its derivatives as anti-HIV- 1 polypeptide drug carriers. Chitooligosaccharides are oligoglucosamine made from 2-20 glucosamine obtained by degrading chitosan by bioengineering technology. It has the important functions of enhancing immunity, reducing blood fat, lowering blood sugar, preventing cancer cell metastasis, and inhibiting cell aging. As a carrier of polypeptide drugs, chitosan oligosaccharide can increase the bioavailability of polypeptide drugs with low absorption rate; Controlling the release of polypeptide drugs; Reduce the irritation to gastrointestinal mucosa; Maintain that stability of polypeptide drugs in vivo; Improve the targeting of drugs.
Genetic engineering bacteria for efficiently expressing human alpha1thymosin, and construction method and application thereof
Project Description: The genetically engineered bacterium is Escherichia coli DH5α, BL2 1(DE3) or BLR(DE3), and its plasmid contains n genes expressing human α 1- thymosin, where n is an integer of 1 ~ 16, and its promoter is IPTG-induced promoter Lac. The pilot study has been completed. Compared with the traditional chemical synthesis method, it has the characteristics of simple process, good activity retention and low cost, and can be widely used for the prevention and treatment of patients with hepatitis B, hepatitis C and cancer.
Development of lysozyme and polypeptide protein products with egg white as raw material
Project Description: Lysozyme is a highly toxic, harmless and safe hydrolytic protease, which is widely used in the fields of food and medicine. Egg white polypeptide is an active polypeptide extracted from egg white, which not only has high nutritional value and low allergenicity, but also is a high-quality protein. Egg white polypeptide can promote the development of children and adolescents and improve their disease resistance. Young women can lose weight by eating it. Young and middle-aged men can resist fatigue and improve energy, and diabetics can help stabilize blood sugar.
High-energy focused ultrasound tumor treatment system
High-energy focused ultrasound therapy system takes advantage of the characteristics of deep penetration, strong directivity and good focusing performance of ultrasound, and uses external focusing technology to converge multiple beams of low-energy ultrasound. Through the coupling effect of water medium, it goes deep into the human tumor to form a high-energy region, resulting in high temperature effect, mechanical effect and cavitation effect of 60- 100 degrees. Causing coagulation necrosis of tumor tissue, that is, losing the ability of focus metastasis. The necrotic "benign" cancer tissue can finally be dissolved and absorbed by the body to achieve the effect of "resection" of the tumor, and the treatment has no effect on the normal tissue outside the target area. From the treatment practice, the technology of high-energy focused ultrasound therapy system is non-invasive and painless, and it can treat malignant tumors accurately and quickly, such as breast cancer and liver cancer. It can also be used to treat benign tumors and other benign diseases, such as hysteromyoma, benign soft tissue tumors, benign breast tumors, etc. In addition, the palliative treatment of residual tumor, tumor recurrence and malignant tumor after routine operation failed, and it was impossible to operate again.
As a non-invasive, painless and accurate method for treating benign and malignant tumor lesions in China, high-energy focused ultrasound tumor treatment system has the following advantages compared with other similar hyperthermia methods for treating tumors at home and abroad:
1. Non-invasive treatment: no surgery, no pain, no trauma, no scar on the epidermis, no radiation side effects, safe and reliable;
2. Standardized treatment: according to the principle of tumor surgical treatment, the diseased tissue in the body is "removed" from the outside, thus making the treatment simple and quick;
3. Conformal therapy: the stereotactic group is suitable for conformal scanning therapy, which ensures that ultrasonic waves penetrate from the outside and destroy the diseased tissues in the body more accurately;
4. Selective treatment: only when the blood vessels less than 2mm in the tumor are destroyed can the safety of treatment be effectively guaranteed by controlling the intensity of ultrasound;
5. Real-time monitoring treatment: the ultrasonic monitoring device controls the whole treatment process and monitors the treatment time in real time;
6. Control the treatment time: after the treatment is completed within the controlled time, the work and life will not be affected;
7. Improve immunity: It can activate the anti-tumor immunity of the host cells, and make them achieve the effect of self-clearing diseased cells and necrotic cells.
The biggest advantage of high-energy focused ultrasound tumor treatment system is that it can avoid local discomfort and systemic adverse reactions caused by chemotherapy and radiotherapy. After high-energy focused ultrasonic scalpel treatment, patients can resume their normal work and life through a short rest.
In order to achieve better comparative study and analysis, we choose the most effective gamma knife, X knife, linear accelerator and neutron knife as the comparison objects, and compare them from the aspects of appearance, performance, curative effect, treatment method, mechanism of action, adaptability, safety and reliability, cost and economic benefit, so as to achieve the best way for people to understand modern tumor treatment.