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Apparatus and method for immunomagnetic cell separation

a technology of immunomagnetic cell and apparatus, applied in the field of magnetic separation device, can solve problems such as difficulty in obtaining sufficient enriched target populations

Inactive Publication Date: 2018-10-11
BIOMAGNETIC SOLUTIONS LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a system and method for magnetic separation of target bioentities from a fluid suspension of target and bystander bioentities using magnetic particles. The system includes a separation chamber with an opening for filling the chamber with a cell suspension and a magnetic element for applying a magnetic field to attract the target bioentities to the collection surface. The technical effects include efficient and effective separation of target bioentities from complex biological mixtures for various applications such as research, diagnostics, and therapeutics.

Problems solved by technology

However, when performing large-scale magnetic separation there are difficulties with obtaining sufficiently enriched target populations.

Method used

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  • Apparatus and method for immunomagnetic cell separation
  • Apparatus and method for immunomagnetic cell separation
  • Apparatus and method for immunomagnetic cell separation

Examples

Experimental program
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Effect test

example 1

The Effect of Gravity on Entrapment of RBC During Magnetic Separation of Target Cells

[0080]For these experiments, CD3+ cells from a CEM cell line (T-lymphoblast, CD3+) were labeled at 108 cells / ml with biotinylated anti-CD3 mouse mAb (Tonbo biotech, San Diego, Calif.), unbound mAb was removed by washing and centrifugation, and cells were magnetically labeled with streptavidin ferrofluid by routine methods. All experiments were done at room temperature. To simulate separations from buffy coats, labeled cells were mixed with fresh bovine erythrocytes in a proprietary buffer. Those RBCs had been recovered from the pellet of centrifuged bovine blood that had been treated with EDTA. Contamination levels of RBC were simulated at 10, 20 and 30% hematocrit. Final volumes for these experiments were 2 ml and separations were done in acrylic cuvettes (1.0×1.0×4.0 cm). From the input numbers of target cells, the geometry of separation and the collection surface area used in these experiments, i...

example 2

The Effect of Gravity on RBC Contamination of CD3+ Cells Recovered from a Buffy Coat

[0086]As noted above, when target cells are isolated from buffy coats, ridding the product of RBCs can require multiple cycles of re-suspension and magnetic separation. To determine whether or not the methods disclosed, specifically separating against gravity and WWOR, are beneficial to the process of recovering CD3+ target cells from a buffy coat, blood of a young normal male was obtained and processed as described in Example 1.

[0087]The buffy coat total white blood cell content was determined by lysing red blood cells in a small buffy coat sample with distilled water and counting the white blood cells in a hemacytometer. The CD3+ cells in the buffy coat were labeled with biotinylated anti-CD3 mouse mAb, unbound mAb removed by centrifugation and cells magnetically labeled with streptavidin ferrofluid by routine methods. Two magnetically labeled cell samples, in duplicate, were separated against grav...

example 3

Isolation of CD3+ Cells from an Apheresis Product

[0091]Starting with an apheresis product of 1010 cells in a volume of about 1.0 L, platelets were initially removed as they are known to interfere with immunomagnetic separations. This can be accomplished by centrifugation or by well-known methods employing membrane technology (e.g., spinning-membrane filtration, Fresenius Kabi AG and Fresenius Kabi USA). The cells were then brought to a volume of approximately 80 ml with an appropriate buffer containing FcR blocking reagents that do not react with common-capture agents, DNAse, protein (e.g., human serum albumin), and other proprietary reagents known to reduce non-specific binding. This mixture was then introduced into the chamber while it was positioned vertically.

[0092]To the 80 ml cell suspension, an anti-CD3-conjugated magnetic particle (for direct labeling) or a common-capture magnetic particle (for indirect labeling) can be added from their respective vessels. Preferably, an ind...

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Abstract

An apparatus and methods are provided for the magnetic separation of target bioentities. The apparatus include a fluid chamber and a magnetic element for drawing target bioentities toward a collection surface of the fluid chamber. The apparatus may include a positioning assembly operable to variable change the position and orientation of the fluid chamber relative to the magnetic element.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims priority under 35 U.S.C. section 119 (e) to U.S. Provisional Application Nos. 62 / 158,845 filed May 8, 2015; 62 / 174,687 filed Jun. 12, 2015; 62 / 213,575 filed Sep. 2, 2015, the entire disclosures of each of the aforementioned applications hereby incorporated by reference.FIELD OF THE INVENTION[0002]The present invention is directed to a magnetic separation device and relates to a system for large-scale magnetic separation of bioentities.BACKGROUND OF THE INVENTION[0003]Isolation of biological materials, including eukaryotic and prokaryotic cells, using magnetic labeling is useful in a variety of research and clinical applications. Magnetically labeled bioentities can be rapidly separated from a heterogeneous population by applying an external magnetic field gradient to an aqueous suspension. However, when performing large-scale magnetic separation there are difficulties with obtaining sufficiently enriched target pop...

Claims

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Application Information

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IPC IPC(8): C12N13/00G01N33/543B03C1/28B03C1/01B03C1/033B03C1/034B01F11/00B01F13/08B01D15/38
CPCC12N13/00G01N33/54326B03C1/288B03C1/01B03C1/0332B03C1/034B01F11/0017B01F13/0818B01D15/3885B03C2201/18B03C2201/22B03C2201/26A61M1/3618B01F31/23B01F33/452C12M47/04
Inventor LIBERTI, PAUL A.KHRISTOV, TODOR R.RITTER, DUSTIN W.
Owner BIOMAGNETIC SOLUTIONS LLC
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