Methods and materials for improved high gradient magnetic separation of biological materials
AssigneeS. Miltenyi
InventorStefan Miltenyi
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Abstract
Improvements in the existing procedures and materials for conduct of high gradient magnetic separation (HGMS) are disclosed. Superior superparamagnetic particles, optionally coated with a polysaccharide or other, usually organic, materials can be prepared in uniform compositions with homogeneous magnetizations. The coating can conveniently be conjugated to a specific binding moiety complementary to a biological material whose purification or separation is desired. In addition, plastic coated matrices which form superior magnetic gradient-intensifying supports are disclosed, along with improved methods and apparatus to conduct HGMS.
Claims17
A method to isolate a biological material from a sample which method comprises
- applying a sample containing the biological material coupled to superparamagnetic particles having a statistical distribution of magnetization when exposed to a magnetic field of sufficient strength to magnetize said particles to more than 90% of their saturation magnetization wherein 90% of said particles in said composition and exposed to said magnetic field have a magnetization within .+-.10% of the mean magnetization of said distribution to a separation chamber which contains a magnetic gradient of sufficient strength to cause retention of the superparamagnetic particles in the chamber,
- allowing the remainder of the sample to pass through the chamber, and
- eluting the superparamagnetic particles from the chamber by introducing into said chamber an eluting fluid containing second superparamagnetic particles of higher saturation magnetization than said first superparamagnetic particles, thereby releasing said biological material from said chamber.
A method to effect separation of at least two components from a mixture, which method comprises
- treating said mixture with a first composition of superparamagnetic particles of a first magnetization having a statistical distribution of magnetization when exposed to a magnetic field of sufficient strength to magnetize said particles to more than 90% of their saturation magnetization wherein 90% of said particles in said composition when exposed to said magnetic field have a magnetization within .+-.10% of the mean magnetization of said distribution, said superparamagnetic particles being conjugated to a specific binding partner complementary to a first component of said mixture and with a second composition of superparamagnetic particles of a second magnetization having a statistical distribution of magnetization when exposed to a magnetic field of sufficient strength to magnetize said particles to more than 90% of their saturation magnetization wherein 90% of said particles in said composition and exposed to said magnetic field have a magnetization within .+-.10% of the mean magnetization of said distribution, said second mean magnetization being different from said first mean magnetization, said superparamagnetic particles being conjugated to a specific binding partner complementary to a second component,
- subjecting the treated mixture to a magnetic gradient in a magnetic separation chamber under conditions wherein at least one of said first and second composition of particles is retained, and
- eluting separate fractions containing said first and second components.
A method to isolate a material from a sample, which method comprises:
- applying a sample containing the material coupled with a superparamagnetic particle to a separation chamber in the presence of a magnetic gradient of sufficient strength to retain the superparamagnetic particles in the chamber, and
- eluting the superparamagnetic particles from the chamber by introduction of a magnetic fluid.
A method to prepare a composition of superparamagnetic colloidal particles of defined magnetic susceptibility which method comprises applying a mixture of superparamagnetic colloidal particles having a range of magnetization to a separation chamber in the presence of a magnetic gradient under conditions wherein at least a fraction of said particles are retained in the chamber, and eluting said retained particles from said chamber by gradual reduction of the magnetic field across the chamber so as to obtain a series of fractions wherein the particles in each fraction having a statistical distribution of magnetization when exposed to a magnetic field of sufficient strength to magnetize said particles to more than 90% of their saturation magnetization wherein 90% of said particles in said composition and exposed to said magnetic field have a magnetization within .+-.40% of the mean magnetization of said distribution.
A method to effect separation of at least two components from a mixture, which method comprises subjecting the mixture to a magnetic gradient in a magnetic separation chamber wherein a first component is coupled to superparamagnetic particles having a first magnetization wherein said particles having a statistical distribution of magnetization when exposed to a magnetic field of sufficient strength to magnetize said particles to more than 90% of their saturation magnetization wherein 90% of said particles in said composition and exposed to said magnetic field have a magnetization within .+-.10% of the mean magnetization of said distribution and a second component is coupled to superparamagnetic particles of a second magnetization wherein said particles having a statistical distribution of magnetization when exposed to a magnetic field of sufficient strength to magnetize said particles to more than 90% of their saturation magnetization wherein 90% of said particles in said composition and exposed to said magnetic field have a magnetization within .+-.10% of the mean magnetization of said distribution, and wherein said second mean magnetization is different from said first mean magnetization
- under conditions wherein at least one of said particles of said first magnetization or second magnetization is retained and
- eluting separate fractions containing said first and second components.
A method to isolate a biological material from a sample which method comprises
- applying a sample containing the biological material coupled to first superparamagnetic particles having a statistical distribution of magnetization when exposed to a magnetic field of sufficient strength to magnetize said particles to more than 90% of their saturation magnetization wherein 90% of said particles in said composition and exposed to said magnetic field have a magnetization within .+-.10% of the mean magnetization of said distribution to a separation chamber which contains a magnetic gradient of sufficient strength to cause retention of the superparamagnetic particles in the chamber,
- allowing the remainder of the sample to pass through the chamber, and
- eluting the superparamagnetic particles from the chamber by reducing the strength of said magnetic field and thereby releasing said biological material from said chamber.
A method to separate biological material using HGMS which method comprises
- applying a sample containing the biological material associated with a particle conjugated to a specific binding partner complementary to the biological material to a separation chamber including a magnetic gradient intensifying matrix in the presence of a magnetic gradient of sufficient strength to cause retention of the magnetized particles in said chamber, and
- eluting the magnetized particles from the chamber;
- wherein said particles are superparamagnetic colloidal particles coated with an organic coating, forming a stable colloid and being prepared by a method which comprises
- subjecting a preparation containing a mixture of superparamagnetic particles coated with an organic coating with a range of magnetization to HGMS under conditions where only a fraction of the particles with the highest magnetization is retained, allowing the remainder of the preparation to flow through said matrix, and eluting the retained particles.
Dependencies are inferred from the claim text.