Superparamagnetic monodisperse particles and method for the production thereof

Abstract

Disclosed herein are monodisperse superparamagnetic beads, having a core-shell structure, and a method for preparing the beads.

Description

technical field

[0001] The present invention relates to monodisperse superparamagnetic microbeads for use in in vitro diagnostic (IVD) assays and other applications, and methods of producing said beads. Background technique

[0002] Monodisperse superparamagnetic microbeads are widely used in the fields of in vitro diagnostics (IVD) and biotechnology. For example, in chemiluminescent assays, monodisperse superparamagnetic microbeads are used to quantitatively detect the amount of antigen or antibody present in a sample by magnetically labeling the antibody or antigen. Monodisperse superparamagnetic microbeads are also a commonly used material in diagnostic kits for nucleic acid (DNA and RNA) extraction prior to polymerase chain reaction (PCR). These beads are easily adapted to automated processes and are used on more than 25,000 routine IVD instruments worldwide. Monodisperse superparamagnetic microbeads can also be used for antibody purification and separation of a variet...

Examples

preparation example Construction

[0191] Therefore, also disclose a kind of preparation method of the monodisperse superparamagnetic microbead with core-shell structure, this method comprises:

[0192] (a) providing monodisperse superparamagnetic precursor microbeads comprising

[0193] The core part, which is formed from a polystyrene polymer matrix material, wherein the polystyrene polymer matrix material encapsulates the first batch of superparamagnetic Fe 3 o 4 Nanoparticles;

[0194] a first shell portion located directly outside the core portion and formed from a cross-linked copolymer matrix material formed from styrene monomers, cross-linking monomers and first functional monomers having functional groups, Among them, the copolymer matrix material encapsulates the second batch of superparamagnetic Fe 3 o 4 nanoparticles; and

[0195] The second housing part, which is located directly outside the first housing part, the second housing part comprises superparamagnetic Fe 3 o 4 Nanoparticle and pol...

Embodiment 1

[0303] Synthesis of Monodisperse Microbeads Bearing -COOH and -OH Groups

[0304] Described below is a "one-pot three-stage" polymerization procedure for the synthesis of monodisperse microbeads bearing -COOH and -OH groups.

[0305] Into a 250 mL three-neck round-bottom glass reactor equipped with a mechanical stirrer, AIBN solution (2 g, 12 wt% in acetone) was added; nitrogen was then introduced from one side neck to remove the acetone solvent. After 10 minutes of nitrogen flow, dry AIBN powder was observed at the bottom of the reactor. Then PVP (0.2 g), ethanol (80 mL) and deionized water (20 mL) were added. The mixture was stirred at room temperature for 5 minutes to obtain a clear solution; the solution was then heated to 60 with an oil bath, followed by the addition of styrene (7.5 mL). After 4 hours a white colloidal solution was produced, indicating polymerization of styrene. The DVB solution (3 mL DVB in 7 mL ethanol) was then added slowly over 30 minutes using a c...

Embodiment 2

[0307] Washing and Surface Modification of Microbeads A-1

[0308] Microbeads of A-1 (50 mL dispersion, mass concentration about 8% by weight) were centrifuged, and then redispersed in ethanol by sonication. The beads were washed with ethanol (50 mL x 2) and a mixture of THF and water (1:1 volume ratio; 50 mL x 2). The obtained microbeads were centrifuged and then redispersed in THF (50 mL) to obtain B-1. SEM images show that these microbeads are spherical and monodisperse, with a diameter of about 1.0 μm.

technical field

[0001] The present invention relates to monodisperse superparamagnetic microbeads for use in in vitro diagnostic (IVD) assays and other applications, and methods of producing said beads. Background technique

[0002] Monodisperse superparamagnetic microbeads are widely used in the fields of in vitro diagnostics (IVD) and biotechnology. For example, in chemiluminescent assays, monodisperse superparamagnetic microbeads are used to quantitatively detect the amount of antigen or antibody present in a sample by magnetically labeling the antibody or antigen. Monodisperse superparamagnetic microbeads are also a commonly used material in diagnostic kits for nucleic acid (DNA and RNA) extraction prior to polymerase chain reaction (PCR). These beads are easily adapted to automated processes and are used on more than 25,000 routine IVD instruments worldwide. Monodisperse superparamagnetic microbeads can also be used for antibody purification and separation of a variet...

Examples

preparation example Construction

[0191] Therefore, also disclose a kind of preparation method of the monodisperse superparamagnetic microbead with core-shell structure, this method comprises:

[0192] (a) providing monodisperse superparamagnetic precursor microbeads comprising

[0193] The core part, which is formed from a polystyrene polymer matrix material, wherein the polystyrene polymer matrix material encapsulates the first batch of superparamagnetic Fe 3 o 4 Nanoparticles;

[0194] a first shell portion located directly outside the core portion and formed from a cross-linked copolymer matrix material formed from styrene monomers, cross-linking monomers and first functional monomers having functional groups, Among them, the copolymer matrix material encapsulates the second batch of superparamagnetic Fe 3 o 4 nanoparticles; and

[0195] The second housing part, which is located directly outside the first housing part, the second housing part comprises superparamagnetic Fe 3 o 4 Nanoparticle and pol...

Embodiment 1

[0303] Synthesis of Monodisperse Microbeads Bearing -COOH and -OH Groups

[0304] Described below is a "one-pot three-stage" polymerization procedure for the synthesis of monodisperse microbeads bearing -COOH and -OH groups.

[0305] Into a 250 mL three-neck round-bottom glass reactor equipped with a mechanical stirrer, AIBN solution (2 g, 12 wt% in acetone) was added; nitrogen was then introduced from one side neck to remove the acetone solvent. After 10 minutes of nitrogen flow, dry AIBN powder was observed at the bottom of the reactor. Then PVP (0.2 g), ethanol (80 mL) and deionized water (20 mL) were added. The mixture was stirred at room temperature for 5 minutes to obtain a clear solution; the solution was then heated to 60 with an oil bath, followed by the addition of styrene (7.5 mL). After 4 hours a white colloidal solution was produced, indicating polymerization of styrene. The DVB solution (3 mL DVB in 7 mL ethanol) was then added slowly over 30 minutes using a c...

Embodiment 2

[0307] Washing and Surface Modification of Microbeads A-1

[0308] Microbeads of A-1 (50 mL dispersion, mass concentration about 8% by weight) were centrifuged, and then redispersed in ethanol by sonication. The beads were washed with ethanol (50 mL x 2) and a mixture of THF and water (1:1 volume ratio; 50 mL x 2). The obtained microbeads were centrifuged and then redispersed in THF (50 mL) to obtain B-1. SEM images show that these microbeads are spherical and monodisperse, with a diameter of about 1.0 μm.

Images