Rensselaer Polytechnic Institute

Other

Bio-rad Laboratories, Inc.

Steven M Cramer

James A Moore

Abhinav A Shukla

Jonathan S Dordick

Yufei Li

Guhan Jayaraman

Sun Kyu Park

Kaushal Rege

Nihal Tugcu

Ranjit R Deshmukh

Amitava Kundu

Hong Chen

Jiali Liao

Shanghui Hu

Russell Frost

James Woo

Khurram M Sunasara

The present disclosure is directed to mixed mode chromatography media comprising a ligand directly attached to a solid support. In some aspects, the ligand has a chemical formula of The mixed mode chromatography media is useful for binding and purifying proteins from a solution.

Mixed mode ligands

Aminoglycoside-polyamines are disclosed along with methods of use thereof in displacement chromatography and as DNA-binding ligands. The aminoglycoside-polyamines are derivatives of carbohydrates, such as sugars, amino sugars, deoxysugars, glycosides, nucleosides and their substituted counterparts. The subject polyamines possess a group in place of at least one hydrogen atom of at least one hydroxyl group of the carbohydrate compound. In these compounds Ris an alkyl group or an azaalkyl group, and Ris a primary or secondary amino group.

Aminoglycoside-polyamine displacers and methods of use in displacement chromatography

A method for separating one or more components of a biomolecule mixture by means of an ion exchange chromatographic system operated in the displacement mode includes sequentially perfusing the system with a first solution including the biomolecule mixture, and a second solution including a displacer having a structure selected from formula I and formula II: 

High-affinity, low molecular-mass displacers for ion-exchange chromotography

A method for separating one or more components of a biomolecule mixture by means of an ion exchange chromatographic system operated in the displacement mode includes sequentially perfusing the system with a first solution including the biomolecule mixture, and a second solution including a displacer having a structure selected from formula I and formula II:

High-affinity, low-molecular-mass displacers for ion-exchange chromatography

An effective technique for the high throughput screening of displacers is described. In this technique, potential displacers are employed to displace a biomolecule (e.g., protein) adsorbed on a chromatographic resin in small-scale batch displacement experiments. The amount of protein displaced from a specific resin by a defined concentration of displacer is determined by monitoring the supermatant for the protein. By evaluating the displaced protein rather than the displacer itself, this technique enables a single detection technique (e.g., absorbance, fluorescence, etc.) to be employed for all batch displacement experiments. By monitoring the amount of protein displaced, the effacy of a large number of potential displacers can be rapidly evaluated. The entire experimental procedure can be carried out rapidly and is thus amenable to high throughput parallel screening of molecules possessing a large range of affinities and physico-chemical properties. The error of the technique is within 5% of protein displaced, thus making it a very reliable technique. The technique can be extended to different stationary phase materials, biomolecules, and modes of interaction.

High throughtput screening of potential displacer molecules

A method for purifying oligonucleotides by displacement chromatography on anion-exchange media, using high affinity, low molecular weight (less than about 10000 Da) displacers, is disclosed. Several examples of high affinity, low molecular weight anionic displacers are provided, including polycyclic aromatic compounds having sulfonic acid moieties attached thereon. The efficacy of the technique for high resolution separation of oligonucleotides is demonstrated for an industrial mixture.

Method for selecting a high affinity, low molecular weight displacer for oligonucleotide purification

High affinity, low molecular weight displacers for oligonucleotide purification

A method for purification of proteins by displacement chromatography in hydrophobic interaction and reversed phase chromatographic systems uses low molecular weight (less than about 10,000) surface-active compounds as displacers. Examples of effective displacers are benzethonium chloride, benzyltributylammonium chloride, and tetrahexylammonium chloride.

Low molecular weight displacers for protein purification in hydrophobic interaction and reversed phase chromatographic systems

A method for the purification of proteins by displacement chromatography on ion exchange media using low molecular weight displacers is disclosed. Several classes of low molecular weight anionic species are exemplified, including aromatic rings having sulfonic acid or carboxylic acid moieties attached thereon, sulfated sugar derivatives, anionic antibiotics, and dendrimeric polymers. Novel compounds useful as displacers are dendrimers of formula ##STR1## wherein R.sup.1 is lower alkyl, n is 2 to 6 and similar dendritic polymers based thereon.

Displacement chromatography of proteins using low molecular weight

A method for the purification of proteins by displacement chromatography on ion exchange media using low molecular weight displacers is disclosed. Several classes of low molecular weight, charged species are exemplified, including aminoacids, peptides, antibiotics and dendrimeric polymers. Novel compounds useful as displacers are dendrimers of formula ##STR1## wherein R.sup.1 is lower alkyl, n is 2 to 6, and X is a common counter anion and similar dendritic polymers based thereon.

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Schenectady, NY, United States of America

Steven M Cramer