141 results about "Size-exclusion chromatography" patented technology

The invention provides a plant exosome as well as an extraction method and application thereof in cosmetics, and belongs to the technical field of exosome extraction. The preparation method of the plant exosome comprises the following steps: mixing plant tissues with a solvent, carrying out high-pressure homogenization treatment, adding immobilized compound enzyme for enzymolysis, filtering, carrying out differential centrifugation on enzymatic hydrolysate, and carrying out size exclusion chromatography and ultrafiltration purification to obtain the plant exosome. Wherein exosome release and protection are considered, and the influence of agglomeration on the exosome extraction effect is avoided; high-pressure homogenization and immobilized compound enzyme enzymolysis are adopted, physical wall breaking is combined with chemical wall breaking, and the yield of the exosome is increased; the immobilized compound enzyme can avoid free enzyme residues, and the subsequent purification step is simplified; the plant exosome can be further purified through size exclusion chromatography and ultrafiltration. The exosome prepared by the method disclosed by the invention can retain the active ingredients of the exosome to the greatest extent, has excellent antioxidant, anti-inflammatory and whitening effects, and is suitable for the field of cosmetics.

Pharmaceutical formulations comprising an antibody that specifically binds to human interleukin-4 receptor alpha (hIL-4Rα) are provided. The formulations may contain, in addition to an anti-IL-4Rα antibody, one or more buffers, at least one amino acid, at least one sugar, and a surfactant. Methods for producing pharmaceutical formulations with reduced lipase activity are also provided, which may include subjecting a drug substance to agitation stress, heat stress, and additional ion exchange or size exclusion chromatography. In one aspect, the pharmaceutical formulations do not have appreciable subvisible particle formation in the presence of lipase, and exhibit a substantial degree of antibody stability during storage and after being subjected to thermal and other physical stresses.

The present invention generally pertains to methods of characterizing crosslinking sites of a protein of interest. In particular, the present invention pertains to the use of size exclusion chromatography, peptide mapping and subunit analysis to identify and quantify crosslinking sites of a protein of interest and determine a contribution of crosslinking to the formation of high molecular weight species.

The invention provides a method for simultaneously detecting the purity of an antibody conjugate and the coupling ratio of a drug antibody based on a size exclusion chromatography method and application of the method. The method comprises the following steps: detecting and analyzing the purity of an ADC (Analog to Digital Converter) sample to be detected and the coupling ratio of the drug antibody by adopting an SEC-UPLC (Single Emission Computation-Ultra Performance Liquid Chromatography) serially connected with an ultraviolet detector; collecting a signal of the sample under the antibody characteristic ultraviolet absorption wavelength and the small molecule characteristic ultraviolet absorption wavelength at the same time; calculating the DAR value and the monomer purity of the ADC molecule according to the chromatographic absorption peak area; a mobile phase adopted by the SEC-UPLC is a sodium chloride-phosphate buffer solution containing 0-10vol% of an organic solvent; the ADC sample to be detected is an ADC molecule coupled with Dxd type small molecules. The invention provides an analysis method which is widely applicable to determination of the DAR value and the monomer purity of the ADC molecule coupled with the Dxd small molecules. The method can simultaneously solve the problems that the DAR value detection and analysis flux of the Dxd small molecules is low and the applicability is limited.

To provide a method and system for protein characterization using online chromatography and electrospray ionization mass spectrometry.SOLUTION: A method for protein characterization comprises the steps of: contacting a sample including protein to a chromatography system with a size exclusion chromatography resin; cleaning the size exclusion chromatography resin using a mobile phase to obtain an eluent containing the protein; and characterizing the protein in the eluent using an electrospray ionization mass spectrometer under non-denaturing conditions.SELECTED DRAWING: Figure 2

The present invention relates to a hinged assembly comprising a polyethylene composition comprising-a high density polyethylene (HDPE) having a density in the range of 950 to 960 kg / m3, measured according to ISO 1183, a melt flow index in the range of 5 to 40 g / 10 min, measured according to ISO 1133 at a load of 2.16 kg and a temperature of 190 DEG C, a weight average molecular weight Mw, measured by size exclusion chromatography, in the range of 50,000 to 110,000 g / mol, a molecular weight distribution Mw / Mn in the range of 4.0 to 10.0; and-a linear low density polyethylene (LLDPE) having a density in the range of 920 to 940 kg / m3, measured according to ISO 1183, a melt flow index in the range of 1 to 20 g / 10 min, measured according to ISO 1133 at a load of 2.16 kg and a temperature of 190 DEG C, a weight average molecular weight Mw, measured by size exclusion chromatography, in the range of 40,000 to 120,000 g / mol, and a molecular weight distribution Mw / Mn in the range of 3 to 7; wherein the polyethylene composition has an overall density, measured according to ISO 1183, in the range of 935 to 950 kg / m3 and an overall melt flow index, measured according to ISO 1133 at a load of 2.16 kg and a temperature of 190 DEG C, of 5 to 30 g / 10 min; wherein the amount of HDPE is in the range of from 30 to 70% by weight of the total amount of HDPE and LLDPE, and wherein the amount of LLDPE is in the range of from 30 to 70% by weight of the total amount of HDPE and LLDPE, and preferably wherein the total amount of HDPE and LLDPE is 100% by weight of the polyethylene composition.

Disclosed herein are methods for characterizing lipid nanoparticles (LNPs) using size-exclusion chromatography. The disclosed methods utilize a denaturing mobile phase (i.e., denaturing SEC) to afford robust disruption of LNPs and to permit the direct injection of LNPs onto a size-exclusion chromatography column. Accordingly, said methods allow for the characterization of formulated LNPs without pre-treatment or other modifications.

The invention discloses an exosome as well as a preparation method and application thereof, and belongs to the technical field of exosomes. The preparation method comprises the following steps: step 1, intermittently immersing and culturing a sterile tissue to obtain a concentrated solution of the exosome I; step 2, collecting the cultured sterile tissues, and performing ultrasonic-assisted extraction to obtain a purified crude extract II; and 3, purifying the concentrated solution obtained in the step 1 and the purified crude extract 2 obtained in the step 2 by size exclusion chromatography to obtain the pure exosome. The exosome obtained by the invention is applied to preparation of cosmetics, health-care foods or medicines for inhibiting skin cell senescence. According to the preparation method disclosed by the invention, impure proteins and non-target particles are effectively removed, the purity of the exosome is improved, the structural integrity of the exosome is kept, false exosome impurities generated by a traditional crushing method are avoided, the yield of the exosome is improved by 3-5 times, and a reliable material basis is provided for subsequent functional research of the exosome.

The present disclosure is directed to stationary phase materials for performing size exclusion chromatography. Embodiments of the present disclosure feature hydroxy-terminated polyethylene glycol surface modified silica particle stationary phase materials, which are optionally also methoxy-terminated polyethylene glycol surface modified.

The invention relates to a chemoenzymatic synthesis method of a high-glucose-type homogeneity mucoprotein core structural domain. The method comprises the following steps: firstly, catalytically synthesizing a key alpha-linked glycosylated amino acid block Fmoc-GalNacalpha-Ser / Thr-OH at a low temperature (-20 DEG C to 0 DEG C); then assembling the building blocks on a repeated polypeptide skeleton rich in proline, serine and threonine at accurate intervals (3-5 amino acid residues) through a solid-phase peptide synthesis technology to form a'sugar chain growth scaffold '; then, a series of high-specificity glycosyl transferases are used for carrying out sequential and directional sugar chain enzymatic extension on the scaffold, and a uniform core structure (such as a core type 3) is constructed; and finally, performing high-efficiency purification by a three-step series method of size exclusion chromatography, ion exchange chromatography and lectin affinity chromatography. The technical bottleneck of inhomogeneity (lt, 70%) of glycoforms in a traditional method is overcome, the mucoprotein core fragment with glycoform homogeneity larger than or equal to 95% and bioactivity highly similar to that of natural mucoprotein can be prepared on a large scale, and the method has wide application prospects in the biomedical fields of mucous membrane protective agents, drug delivery carriers and the like.

This disclosure relates to methods for performing size exclusion chromatography. Embodiments of this disclosure are characterized by methods for improving the separation of protein analytes in size exclusion chromatography, for example, by using a low concentration of amino acids or derivatives thereof in the mobile phase and using a stationary phase comprising porous particles with surface-modified polyethylene groups capped with hydroxyl groups.

The present application relates to systems and methods for purifying polypeptides, such as His tagged polypeptides, using immobilized metal chelate affinity chromatography (IMAC). The method comprises loading a composition comprising the polypeptide onto an IMAC matrix; eluting the polypeptide from the first IMAC matrix; performing buffer exchange to remove the elution buffer from the first eluate; loading the first eluate from the buffer exchange onto a second IMAC substrate; eluting the polypeptide from the second IMAC matrix; and optionally purifying the eluate from the second IMAC matrix using size exclusion chromatography. In some aspects, the method is fully automated.

The invention relates to the technical field of cell separation and extraction, and provides a method for extracting extracellular vesicles from induced sputum. Comprising the following steps: S1, atomizing and collecting induced sputum, picking out sputum plugs, and adding an isotonic phosphate buffer solution which contains 0.1 wt% of drocisteine and 0.1 wt% of tween-20 and has the pH value of 6.0 for dissolving, so as to obtain an induced sputum dissolving solution rich in extracellular vesicles; wherein the dissolving temperature is 37 + / -0.1 DEG C, and digestion is performed for 15 + / -1 min under the condition that the rotating speed of a shaking table is 50 + / -1 rpm; and S2, extracting the extracellular vesicles from the induced sputum dissolving solution through a negative pressure size exclusion chromatography method. According to the invention, the multi-component dissolving agent of the hydroxymethylsteine and the Tween-20 is adopted to promote the dissolving of sputum and protect protein and nucleic acid in extracellular vesicles from being degraded; the negative pressure size exclusion chromatography can accurately control the flow velocity driven by negative pressure, ensures stable migration of extracellular vesicles in a chromatographic column, and avoids vesicle rupture caused by too fast flow velocity or low separation efficiency caused by too slow flow velocity.

The present disclosure is directed to methods of performing direct affinity-size exclusion chromatography, wherein there is a direct elution of the sample from the affinity chromatography column to the SEC column. The methods described herein allow for rapid and robust purification of target analytes from heterogeneous samples, and mitigate the need for complicated valve switching, buffer exchange, or other sample manipulation.

To provide a simple, high-resolution method of isolating and quantifying proteins, in particular antibody drugs and aggregates thereof, and suspended nanoparticles using the SEC (Size Exclusion Chromatography) method.SOLUTION: A size exclusion chromatography method is provided, involving analyzing nanoparticles with diameters in a range of 10 nm to 150 nm and proteins in an aqueous eluent containing one or more non-ionic surfactants.SELECTED DRAWING: Figure 11

The invention discloses a method for separating and purifying protein from Tianshan red deer wrinkled stomach.The method comprises the steps that the Tianshan red deer wrinkled stomachs serve as research objects, adipose tissue and connective tissue of the fresh Tianshan red deer wrinkled stomachs are stripped, and then drying and degreasing treatment is conducted; the method comprises the following steps: extracting, freezing and thawing and centrifuging active proteins by using a freezing and thawing method, dialyzing supernate, freezing and drying to obtain crude protein powder, and purifying by using an ultrafiltration tube, HW-55F size exclusion chromatography and C8 reverse chromatography in combination with high performance liquid chromatography to finally obtain three high-purity Tianshan red deer wrinkled stomach proteins with anti-tumor activity, which are respectively named as S1, S2 and S3. The yields are respectively 1.10%, 1.11% and 0.61%, and the protein contents are 87.14% + / -2.16%, 84.65% + / -3.29% and 82.06% + / -1.25%. The IC50 of the inhibitory activity of the three proteins on HCT-116 colon cancer cells is respectively 55.89 [mu] g / mL + / -2.97, 4.44 [mu] g / mL + / -0.23 and 11.31 [mu] g / mL + / -0.75.

The application discloses a purification method of plant exosome, and belongs to the technical field of biological separation. The method comprises the following steps: obtaining plant juice or tissue extract, pre-treating the plant juice or tissue extract through coarse filtration and centrifugation, incubating the plant juice or tissue extract with magnetic microspheres modified with carboxyl or epoxy groups in a buffer containing calcium ions, capturing the complex by using an external magnetic field, washing the complex, eluting the complex by using a competitive elution buffer containing N-acetylglucosamine, and further purifying the complex by using a size exclusion chromatography column to obtain the plant exosome. The exosome is specifically captured by the magnetic microspheres, and the double purification strategies of competitive elution and size exclusion chromatography are combined, so that the recovery rate and purity of the exosome are significantly improved. The method is simple in operation, short in time consumption, good in repeatability, and suitable for the large-scale preparation of exosomes of various plant sources. The plant-derived exosome prepared by the method can be applied to the fields of drug delivery systems, cosmetics, respiratory health and sensory health product development.

The present disclosure generally pertains to methods for identifying the binding of a peptide or protein to a binding partner using affinity-resolved size exclusion chromatography coupled to mass spectrometry.

The invention provides an extraction and freeze-drying preservation technology of stem cell exosomes. The extraction and freeze-drying preservation technology comprises the following steps: (1) primarily concentrating the exosomes by adopting an ultrafiltration method; (2) further purifying by combining size exclusion chromatography (SEC), and removing impurity proteins and non-exosome vesicles; (3) a trehalose-nanogold composite protective agent is adopted for freeze-drying treatment, so that the structural stability of the exosome in the freeze-drying process is improved; and (4) optimizing a freeze-drying process, so that the exosome can be stably preserved for 12 months at room temperature. According to the method, the recovery rate and the biological activity of the exosome are remarkably improved, and the method is suitable for large-scale production and clinical application.

The invention discloses a method for purifying plant exosomes, which belongs to the technical field of biological separation and comprises the following steps: acquiring plant juice or tissue extracting solution, performing rough filtration and centrifugal pretreatment, incubating the plant juice or tissue extracting solution and magnetic microspheres of which the surfaces are modified with carboxyl groups or epoxy groups in a buffer solution containing calcium ions, capturing a compound by using an external magnetic field, and purifying the plant exosomes. After cleaning, eluting by adopting a competitive elution buffer solution containing N-acetylglucosamine, and finally further purifying by virtue of a size exclusion chromatographic column, so as to obtain the plant exosome. The exosome is specifically captured through the magnetic microspheres, the recovery rate and purity of the exosome are remarkably improved by combining a dual purification strategy of competitive elution and size exclusion chromatography, operation is easy and convenient, consumed time is short, repeatability is good, and the method is suitable for large-scale preparation of exosomes of various plant sources; the plant-derived exosome prepared by the method can be applied to the fields of product development of drug delivery systems, functional foods, cosmetics, respiratory health and sensory health and the like.

A method of separating free target analyte from protein-bound target analyte is described. Such can include obtaining an aqueous sample containing a target analyte in a free form (free target analyte) and the target analyte in a protein-bound form (protein-bound target analyte), passing the aqueous sample through a size exclusion chromatography matrix with a molecular weight cut off sufficient to allow the free target analyte to permeate into pores of the size exclusion chromatography matrix and exclude the protein-bound target analyte, whereupon the free target analyte adheres to and is immobilized by the size exclusion chromatography matrix and the protein-bound target analyte does not adhere to by the size exclusion chromatography matrix, separating the free target analyte from the protein-bound analyte by removing the protein-bound target analyte from the size exclusion chromatography matrix, and eluting the free target analyte from the size exclusion chromatography matrix with an organic solvent.

A method is provided to simultaneously identify and quantify critical structure attributes (CSAs) in proteoforms or proteoform families that includes the steps of:1. injecting a sample into a reagent stream of one or more detection enhanced molecular recognition reagents;2. incubating the combined sample:reagent(s) during migration through an analytical platform the causes molecular recognition reagent(s) to be sequestered by a specific critical structure attribute (CSA) in the analyte to form a luminon (An:*Sas) complex;3. using a sequestron selector in the reagent stream to overtake the luminon complex in a down-stream size exclusion chromatography (SEC) column to achieve mixing and formation of an Nc˜Pas:An:*Sas, Pas:An:*Sas, or *Pas:An:*Sas sequestome complex;4. resolving this sequestome complex from any unbound affinity selector(s) such as *Sas or *Pas, and non-analytes in the SEC column;5. transporting the resolved sequestome complex into a flow-through detection means that detects and quantifies CSAs in fluorescent labeled *Sas or *Pas bearing luminon or sequestome complex to generate data for the construction of critical structure attribute ratio plots as a function of elapsed fermentation time.

Disclosed is a column comprising particles having an average particle size ranging from about 1 [mu] m to about 5 [mu] m; the particles have an average pore size ranging from about 450 to about 3000; and the particles have an average pore volume ranging from about 0.1 cm < 3 > / g to about 5 cm < 3 > / g. The column may be a size exclusion chromatography column. The column can be used in a method for separating a monomer from a viral analyte, and the method is also disclosed.

The invention belongs to the technical field of starch structure regulation and function modification, and provides a preparation method of heat-resistant and digestion-resistant starch microspheres. The method comprises the following steps: by taking high amylose as a raw material, carrying out acidolysis and debranching treatment to obtain an amylose mixed component, and further carrying out accurate grading on the amylose according to the molecular size through volume exclusion chromatography to obtain an amylose fraction with narrow polymerization degree distribution and uniform chain length. Then recrystallization induction is carried out on the fractions, autoclaving treatment is carried out under the condition that the water content is controlled, amylose molecules are subjected to ordered rearrangement, and the starch microspheres which are compact in structure and high in crystallinity are constructed. The prepared resistant starch microspheres have excellent thermal stability and enzymolysis resistance, and still keep higher resistant starch content after being boiled. The method realizes accurate regulation and control of the chain length of the amylose and synergistic improvement of heat resistance and digestion resistance, is suitable for the fields of functional food and nutrition and health, and has a good application prospect.

Provided is a novel porous cellulose medium that can efficiently separate a large target molecule in a calibration standard. A porous cellulose medium including a porous cellulose particle having a particle size from 1 to 600 μm, wherein, in sieving the porous cellulose medium for classification and using a fraction corresponding to aperture openings between 53 μm and 106 μm as a support for size exclusion chromatography, a polyethylene oxide standard is run through size exclusion chromatography with pure water as a mobile phase, and a weight average molecular weight Mw and a gel partition coefficient Kav of the polyethylene oxide standard satisfy Relationships (A) and (B) above:in a case where 4.80≤log Mw≤5.50,Kav>−0.445×log Mw+2.55  (A)in a case where 5.75≤log Mw,0≤Kav<0.19  (B).

The present invention relates to a process for isolating Small Extracellular Vesicles secreted by umbilical cord blood mononuclear cells (UCBMNCs) and compositions comprising said Small Extracellular Vesicles, which are useful to be applied to autoimmune diseases therapeutics or prophylactics and / or cosmetic purposes.The proposed process for isolating UCBMNCs Small Extracellular Vesicles comprises three main steps: i) a first step of sequential centrifugation, ii) a second step of microfiltration combined with ultrafiltration (UF), and iii) a third step of size exclusion chromatography (SEC) and aims to achieve highly pure Small Extracellular Vesicles and in a higher yield.The SEVs compositions comprise specific type of proteins, RNA and lipids, that enables them to be very effective when applied to inflammatory and autoimmune diseases therapeutics, such as psoriasis, lupus, atopic dermatitis, eczema, etc. and also to cosmetic or prophylactic compositions.Therefore, the present invention lays in the technical domain of pharmaceuticals, medicine, cosmetics, research and development in cellular biology and appliances thereof.

The present invention provides a lotus-derived extracellular vesicle (LDEV), which are extracted from lotus leaves by a group selected from the following extraction methods: polymer precipitation method, ultracentrifugation method, ultrafiltration method, density gradient centrifugation method and size-exclusion chromatography. The present invention also provides an anti-inflammatory composition containing the LDEV and the use of the LDEV. The LDEV extracted through different separation methods in the present invention have similar particle sizes and stable zeta potentials. In addition, the present invention has experimentally confirmed that the LDEV can be used to alleviate inflammatory reactions or wound healing, and can further be prepare as anti-inflammatory or wound healing drugs, compositions or nutritional supplements.

Described herein are systems and methods used for carrying out a two-dimensional liquid chromatography process using size exclusion chromatography as a first dimension.

The disclosure relates to methods for purifying RNA from a sample using size exclusion chromatography (SEC). Uses of a SEC medium for purifying RNA from a sample are also provided.

The present disclosure is directed to methods for characterization of a sample by size exclusion chromatography (SEC), the sample including intact lipid nanoparticles (LNPs). The method generally includes loading the sample on a chromatographic column having an SEC packing material disposed therein, flowing a mobile phase through the SEC packing material to elute the intact LNPs, and detecting the eluted intact LNPs. The mobile phase includes an organic solvent, an aqueous buffer, and a non-ionic surfactant.