The present invention is directed to a method of purifying oligonucleotides using hydrophobic interaction chromatography.

A liquid chromatography packing material which includes a polymer packing material into which 1.50 mmol or more of carboxyl groups are introduced per 1 g of the packing material, and an index indicating pH of the surface of the polymer packing material as determined by hydrophilic interaction chromatography (HILIC) is 1.30 or more and the index indicating hydrophilicity of the surface of the polymer packing material as determined by a hydrophilic interaction chromatography (HILIC) is from 1.00 to 1.30.

The present invention relates to a method of separating and analyzing a mixture of oligonucleotides, including performing liquid chromatography using a column packed with a packing material obtained by fixing a diol to a surface of each of porous particles formed of a crosslinked organic polymer. According to this method, the oligonucleotides can be separated and analyzed with higher sensitivity compared to cases where columns having silica gel as a base material are used. In addition, the column can be washed with an alkaline solution.

In one aspect, the present invention provides a chromatographic stationary phase material for various different modes of chromatography represented by Formula 1: [X](W).sub.a(Q).sub.b(T).sub.c (Formula 1). X can be a high purity chromatographic core composition having a surface comprising a silica core material, metal oxide core material, an inorganic-organic hybrid material or a group of block copolymers thereof. W can be absent and/or can include hydrogen and/or can include a hydroxyl on the surface of X. Q can be a functional group that minimizes retention variation over time (drift) under chromatographic conditions utilizing low water concentrations. T can include one or more hydrophilic, polar, ionizable, and/or charged functional groups that chromatographically interact with the analyte. Additionally, b and c can be positive numbers, with the ratio 0.05≤(b/c)≤100, and a≥0.

A method of enhancing a mass spectral signal is disclosed. The method can include contacting a sample to a separation column under conditions that permit sample components to bind to the substrate; applying a first mobile gradient to the separation column, wherein the first mobile phase gradient comprises trifluoroacetic acid (TFA) and a small molecule additive (e.g., an amino acid) or formic acid (FA) and a small molecule additive (e.g., an amino acid); applying a second mobile gradient to the separation column, wherein the second mobile phase gradient comprises TFA in acetonitrile (ACN) and a small molecule additive (e.g., an amino acid) or formic acid (FA) in ACN and a small molecule additive (e.g., an amino acid); and performing mass spectrometric analysis on eluted sample components.

The invention relates to poly-amide bonded hydrophilic interaction chromatography (HILIC) stationary phases and novel HILIC methods for use in the characterization of large biological molecules modified with polar groups, known to those skilled in the art as glycans. The invention particularly provides novel, poly-amide bonded materials designed for efficient separation of large biomolecules, e.g. materials having a large percentage of larger pores (i.e. wide pores). Furthermore, the invention advantageously provides novel HILIC methods that can be used in combination with the stationary phase materials described herein to effectively separate protein and peptide glycoforms by eliminating previously unsolved problems, such as on-column aggregation of protein samples, low sensitivity of chromatographic detection of the glycan moieties, and low resolution of peaks due to restricted pore diffusion and long intra/inter-particle diffusion distances.

By using a graft polymer comprising a dendritic polymer with a styrene skeleton and a hydrophilic polymer grafted to a terminal thereof, a temperature-responsive substrate for cell culture having a temperature-responsive surface for cell culture that allows cells to be cultured with high efficiency and which yet allows cultured cells to be exfoliated in a short period of time and with high efficiency by simply changing the temperature of the substrate surface can be prepared conveniently. If this temperature-responsive substrate for cell culture is used, cells obtained from a variety of tissues can be cultured with high efficiency. If this culture method is utilized, cultured cells can be exfoliated intact in a short amount of time with high efficiency. In addition, by using this graft polymer, a wide range of peptides and proteins can also be separated by simply changing the temperature of a chromatographic carrier. This allows for convenient separation procedure and improves the efficiency of separating operations. What is more, the stereoregularity of the dendritic polymer per se may be utilized to enable separation of solutes based on differences in their molecular structures.

A method for the manufacture of highly purified .sup.68Ge material for radiopharmaceutical purposes. The invention particularly concerns the production of .sup.68Ge-API (API=Active Pharmaceutical Ingredient) solution complying with the Guidelines for good manufacturing practices (GMP). Starting material for the method of the present invention can be a .sup.68Ge stock solution of commercial or other origin as raw material. Such .sup.68Ge containing raw solutions are purified from potential metal and organic impurities originating from production processes. The radiochemical method disclosed is based on a twofold separation of .sup.68Ge from organic and metallic impurities with two different adsorbent materials. During the first separation phase .sup.68Ge is purified from both organic and metallic impurities by adsorption in germanium tetrachloride form, after which hydrolyzed .sup.68Ge is purified from remaining metallic impurities by cation exchange. The final .sup.68Ge-API-product e.g. fulfills the regulatory requirements for specifications of the GMP production of .sup.68Ge/.sup.68Ga generators.

Methods of purifying targeted oligonucleotides within a reaction mixture using hydrophilic interaction liquid chromatography (HILIC) is disclosed. One of the methods in accordance with the present disclosure includes screening the targeted oligonucleotides within the reaction mixture with HILIC to create an initial reaction mixture profile; determining an elution percentage for the targeted oligonucleotides; focusing a HILIC elution gradient around the elution percentage of the targeted oligonucleotides; and purifying the targeted oligonucleotides with HILIC using the focused elution gradient at room temperature. Some embodiments can utilize mass triggering for fraction collection of the targeted oligonucleotides. Some embodiments can utilize UV triggering when the mass falls outside of the mass range of the MS detector.

The invention relates to a method for controlling preparative liquid chromatography, comprising the following steps, at least a part of said steps being implemented by a computer comprising a processor and a display screen coupled to said processor: (a) selecting an analytical liquid chromatography method from among thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), (b) inputting analytical liquid chromatography data obtained by the method selected at step (a) for a product to be purified, (c) accessing a table of separating tools available to the user to implement said preparative liquid chromatography, (d) from said analytical liquid chromatography data and table of available separating tools, selecting an optimal separating tool from said table and computing preparative liquid chromatography operating conditions for said selected separating tool.