Methods and materials for high gradient magnetic separation of biological materials

A mechanically stiff coated columnar matrix for locally intensifying a magnetic gradient in a separation chamber of an HGMS apparatus in the conduct of HGMS (High Gradient Magnetic Separation), said coated columnar matrix comprising magnetically susceptible particles comprising surfaces coated with a solid polymer coating and arranged in a column;

• wherein said matrix further comprises interstices which let liquids pass and is magnetized in the presence of a magnetic field,
• said matrix having been coated using a method comprising:
• treating said magnetically susceptible particles with a coat-forming liquid preparation in the separation chamber, thereby covering said surfaces of said magnetically susceptible particles, said preparation comprising a monomer capable of forming a polymer or a prepolymerized polymer;
• removing excess coat-forming liquid from said magnetically susceptible particles;
• and
• effecting the formation of said solid polymer coating to form said coated matrix,
• said solid polymer coating being essentially impermeable to ions and comprising less than 10 percent water by weight based on a total weight of said coating.

A method of fractionating biological material from a sample in a separation chamber using a coated matrix for intensifying locally a magnetic gradient in a separation chamber in the conduct of HGMS (High Gradient Magnetic Separation), wherein the separation chamber includes a separation vessel, and the coated matrix, prior to coating, includes an uncoated matrix comprising magnetically susceptible beads comprising surfaces;

• wherein the coated matrix has interstices which let liquids pass and wherein the coated matrix is magnetized in the presence of a magnetic field;
• wherein the matrix has been coated in a preparation vessel using a method comprising:
• treating the uncoated matrix with a coat-forming liquid preparation in the preparation vessel, thereby covering the surfaces of the magnetically susceptible beads, wherein the preparation comprises a monomer capable of forming a polymer or a prepolymerized polymer;
• removing excess coat-forming liquid from the so covered matrix;
• and
• effecting the formation of a solid polymer coating of the uncoated matrix to form the coated matrix, the solid polymer coating being essentially impermeable to ions and comprising less than 10% water by weight based on a total weight of the coating;
• transferring the coated matrix from the preparation vessel to the separation vessel;
• said method of fractionating comprising:
• magnetizing said coated matrix by applying a magnetic field and applying said sample comprising the biological material bound by superparamagnetic particles to said chamber, said magnetizing being of sufficient strength to retain a part of said sample in said chamber;
• allowing a remainder of said sample to pass through said chamber;
• and
• eluting said part of said sample retained in said chamber from said chamber.

A method of fractionating biological material from a sample in a separation chamber using a coated matrix for intensifying locally a magnetic gradient in a separation chamber in the conduct of HGMS (High Gradient Magnetic Separation), wherein the separation chamber includes a separation vessel, wherein the coated matrix, prior to coating, comprises an uncoated matrix of magnetically susceptible beads comprising surfaces, and wherein the coated matrix has been coated using a method comprising:

• treating the uncoated matrix with a coat-forming liquid preparation in the separation vessel, thereby covering the surfaces of the magnetically susceptible beads, wherein the preparation comprises a monomer capable of forming a polymer or a prepolymerized polymer;
• removing excess coat-forming liquid from the beads;
• and
• effecting the formation of a solid polymer coating to form the coated matrix, wherein the solid polymer coating is essentially impermeable to ions and comprises less than about 10% water by weight based on a total weight of the coating;
• wherein the coated matrix includes interstices which let liquids pass and wherein the coated matrix becomes magnetized in the presence of a magnetic field;
• said method of fractionating comprising:
• magnetizing said coated matrix by applying, to the separation chamber, a magnetic field and a sample comprising biological material bound by superparamagnetic particles to be separated, the magnetization being of sufficient strength to retain a part of said sample in said chamber;
• allowing the remainder of said sample to pass through said chamber;
• and
• eluting said part of said sample retained in said chamber from said chamber.

An article of manufacture for use in a high gradient magnetic separation of biological material, comprising:

• a) a matrix for intensifying a magnetic gradient in a separation chamber of a high gradient magnetic separation apparatus, wherein said matrix comprises:
• i) a magnetically susceptible metallic material which is capable of being magnetized in the presence of a magnetic field;
• and
• ii) a solid polymer coating on said metallic material, wherein said polymer coating is impermeable to ions and comprises less than 10% water by weight based on a total weight of said polymer coating on said metallic material;
• and
• b) a composition of superparamagnetic colloidal particles dimensioned to pass through the matrix, wherein said particles
• i) comprise an organic coating to provide functional groups for conjugation to a specific binding moiety;
• ii) are conjugated to said specific binding moiety which is complementary to a subset of the biological material, via said organic coating;
• and
• iii) comprise a lower size limited to 0.01 .mu.m and an upper size limit of 0.2 .mu.m .

An article of manufacture for use in a high gradient magnetic separation of biological material, comprising:

• a) a matrix for intensifying a magnetic gradient in a separation chamber of a high gradient magnetic separation apparatus, wherein said matrix comprises:
• i) a magnetically susceptible metallic material which is capable of being magnetized in the presence of a magnetic field;
• and
• ii) a solid polymer coating on said metallic material, wherein said polymer coating comprises less than 10% water by weight based on a total weight of said polymer coating on said metallic material;
• and
• b) a composition of superparamagnetic colloidal particles dimensioned to pass through the matrix, wherein said particles
• i) comprise an organic coating to provide functional groups for conjugation to a specific binding moiety;
• ii) are conjugated to said specific binding moiety which is complementary to a subset of the biological material, via said organic coating;
• and
• iii) comprise a lower size limited to 0.04 .mu.m and an upper size limit of 0.1 .mu.m.

An article of manufacture for use in a high gradient magnetic separation of biological material, comprising:

• a) a matrix for intensifying a magnetic gradient in a separation chamber of a high gradient magnetic separation apparatus, wherein said matrix comprises:
• i) a magnetically susceptible metallic material which is capable of being magnetized in the presence of a magnetic field;
• and
• ii) a solid polymer coating on said metallic material, wherein said polymer coating comprises less than 10% water by weight based on a total weight of said polymer coating on said metallic material;
• and
• b) a composition of superparamagnetic colloidal particles dimensioned to pass through the matrix, wherein said particles
• i) comprise an organic coating to provide functional groups for conjugation to a specific binding moiety;
• ii) are conjugated to said specific binding moiety which is complementary to a subset of the biological material, via said organic coating;
• and
• iii) comprise a lower size limited to 0.01 .mu.m and an upper size limit of 0.2 .mu.m.

A mechanically stiff coated columnar matrix for locally intensifying a magnetic gradient in a separation chamber of an HGMS apparatus in the conduct of HGMS (High Gradient Magnetic Separation), said separation chamber including a separation vessel, said columnar matrix comprising magnetizable particles and interstices which let liquid pass and said columnar matrix being magnetized in the presence of a magnetic field;

• said columnar matrix having been coated in a preparation vessel using a method including the steps of
• treating said matrix with a coat-forming liquid preparation in a preparation vessel, thereby covering the surfaces of said matrix of magnetizable particles, wherein the preparation comprises a monomer capable of forming a polymer or a prepolymerized polymer;
• removing excess coat-forming liquid from said so covered matrix;
• and
• effecting the formation of a solid polymer coating of said matrix to form a solid, coated columnar matrix, said solid polymer coating containing less than about 10 percent water by weight based on the total weight of the solid coating;
• and
• transferring said columnar matrix from said preparation vessel to said separation chamber of said apparatus.

A mechanically stiff coated columnar matrix for locally intensifying a magnetic gradient in a separation chamber of an HGMS apparatus in the conduct of HGMS (High Gradient Magnetic Separation), said coated columnar matrix including a matrix comprised of finely divided magnetically susceptible particles coated with a solid polymer coating and arranged in a column;

• wherein said coated matrix has interstices which let liquids pass and is magnetized in the presence of a magnetic field,
• said coated matrix having been coated using a method including the steps of
• treating said matrix with a coat-forming liquid preparation in said separation vessel, thereby covering the surfaces of said matrix of magnetizable particles, said preparation comprising a monomer capable of forming a polymer or a prepolymerized polymer;
• removing excess coat-forming liquid from said so covered matrix;
• and
• effecting the formation of said solid polymer coating to form said coated matrix, said solid polymer coating containing less about than 10 percent water by weight based on the total weight of said coating.

A method for fractionating biological material from a liquid in a separation chamber using a magnetically stiff coated columnar matrix for locally intensifying a magnetic gradient in a separation chamber of an HGMS apparatus in the conduct of HGMS (High Gradient Magnetic Separation), the separation chamber comprising a separation vessel, the columnar matrix comprising magnetizable particles comprising surfaces and interstices which let liquid pass and the columnar matrix being magnetized in the presence of a magnetic field;

• the matrix having been coated in a preparation vessel using a method comprising:
• treating the magnetizable particles with a coat-forming liquid preparation in the preparation vessel, thereby covering the surfaces of the matrix of magnetizable particles, which preparation comprises a monomer capable of forming a polymer or a prepolymerized polymer;
• removing excess coat-forming liquid from the magnetizable particles;
• and
• effecting the formation of a solid polymer coating of the magnetizable particles to form the columnar matrix, the solid polymer coating being essentially impermeable to ions and comprising less than 10 percent water by weight based on a total weight of the coating;
• and
• transferring the columnar matrix from the preparation vessel to the separation chamber of the apparatus;
• said method for fractionating comprising:
• magnetizing the coated columnar matrix by applying a magnetic field and applying a sample comprising biological material bound by superparamagnetic particles to be separated to the chamber, said magnetizing being of sufficient strength to retain a part of the sample in the chamber,
• allowing the remainder of the sample to pass through the chamber;
• and
• eluting the part of the sample retained in the chamber from the chamber.

A method of fractionating biological material from a liquid in a separation chamber using a mechanically stiff coated columnar matrix for locally intensifying a magnetic gradient in a separation chamber of an HGMS apparatus in the conduct of HGMS (High Gradient Magnetic Separation), the separation chamber comprising a separation vessel, the columnar matrix comprising magnetizable particles and interstices which let liquid pass and the columnar matrix being magnetized in the presence of a magnetic field;

• the columnar matrix having been coated in a preparation vessel using a method comprising:
• treating the matrix with a coat-forming liquid preparation in a preparation vessel, thereby covering the surfaces of the matrix of magnetizable particles, wherein the preparation comprises a monomer capable of forming a polymer or a prepolymerized polymer;
• removing excess coat-forming liquid from the so covered matrix;
• and
• effecting the formation of a solid polymer coating of the matrix to form a solid, coated columnar matrix, the solid polymer coating comprising less than about 10 percent water by weight based on the total weight of the solid coating;
• and
• transferring the columnar matrix from the preparation vessel to the separation chamber of the apparatus;
• said method of fractionating comprising:
• magnetizing the coated columnar matrix by applying a magnetic field and applying a sample comprising biological material bound by superparamagnetic particles to be separated to the chamber, said magnetizing being of sufficient strength to retain a part of the sample in the chamber;
• allowing the remainder of the sample to pass through the chamber;
• and
• eluting the part of the sample retained in the chamber from the chamber.

A method of fractionating biological material from a liquid in a separation chamber using a mechanically stiff coated matrix for locally intensifying a magnetic gradient in a separation chamber of an HGMS apparatus in the conduct of HGMS (High Gradient Magnetic Separation), the coated columnar matrix comprising a matrix comprised of finely divided magnetically susceptible particles coated with a solid polymer coating and arranged in a column;

• wherein the coated matrix comprises interstices which let liquids pass and is magnetized in the presence of a magnetic field,
• the coated matrix having been coated using a method comprising:
• treating the matrix with a coat-forming liquid preparation in the separation vessel, thereby covering the surfaces of the matrix of magnetizable particles, the preparation comprising a monomer capable of forming a polymer or a prepolymerized polymer;
• removing excess coat-forming liquid from the so covered matrix;
• and
• effecting the formation of the solid polymer coating to form the coated matrix, the solid polymer coating comprising less about than 10 percent water by weight based on the total weight of the coating;
• said method of fractionating comprising:
• magnetizing the coated columnar matrix by applying a magnetic field and applying a sample comprising biological material bound by superparamagnetic particles to be separated to the chamber, said magnetizing being of sufficient strength to retain a part of the sample in the chamber;
• allowing the remainder of the sample to pass through the chamber;
• and
• eluting the part of the sample retained in the chamber from the chamber.