IP Library › Granted Patent US 12,332,239
Granted Patent B2
US 12,332,239 · App. 18/139,242 · Granted Jun 17, 2025

Magnetic removal or identification of damaged or compromised cells or cellular structures

Inventor: Kristie Krug (Upper Arlington, OH)
Assignee: INGURAN, LLC
G01N33/5434B03C1/01B03C1/015B03C1/02B03C1/023B03C1/288C12N13/00G01N33/54333G01N33/54346G01N33/54353G01N33/56966B03C2201/18B03C2201/26
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Quick Facts
Patent No.
US 12,332,239
App. No.
18/139,242
Granted
Jun 17, 2025
Kind
B2
Abstract

A method for magnetic cellular manipulation may include contacting a composition with a biological sample to form a mixture. The composition may include a plurality of particles. Each particle in the plurality of particles may include a magnetic substrate. The magnetic substrate may be characterized by a magnetic susceptibility greater than zero. The composition may also include a chargeable silicon-containing compound. The chargeable silicon-containing compound may coat at least a portion of the magnetic substrate. The biological sample may include cells and/or cellular structures. The method may also include applying a magnetic field to the mixture to manipulate the composition.

Claims (7)

1. A method for processing, the method comprising:

a) contacting sperm cells with a plurality of magnetic particles, the plurality of magnetic particles having an average hydrodynamic diameter of 300 nm and each magnetic particle in the plurality of magnetic particles comprising a negative zeta potential charge and a chargeable silicon-containing compound, the magnetic particles binding to the sperm cells through an electrical charge interaction; and

b) applying a magnetic field to the plurality of magnetic particles.

2. The method of claim 1 , wherein each magnetic particle comprises Fe 3 O 4 .

3. The method of claim 1 , wherein the chargeable silicon-containing compound comprises 2-(carbomethoxy)ethyltrimethoxysilane.

4. The method of claim 1 , further comprising the step of diluting the sperm cells with a buffered media prior to step a).

5. The method of claim 1 , wherein each magnetic particle in the plurality of magnetic particles is between 30 nm to 1000 nm in size.

Assignments (4)
SECURITY INTEREST Recorded Nov 24, 2025
From: INGURAN, LLC
To: BANK OF AMERICA, N.A., AS ADMINISTRATIVE AGENT
Reel/Frame 073783/0924 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 12, 2023
From: KRISTIE, KRUG MARIE
To: CYTOLUTIONS, LLC
Reel/Frame 063622/0637 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 12, 2023
From: CYTOLUTIONS, LLC
To: CELLULAR PROTECTION TECHNOLOGIES, LLC
Reel/Frame 063622/0686 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 12, 2023
From: CELLULAR PROTECTION TECHNOLOGIES, LLC
To: INGURAN, LLC
Reel/Frame 063622/0693 →
Continuity (5)
Continuation 16438215 · Jun 11, 2019
Continuation 15713391 · Sep 22, 2017
Continuation 13974139 · Aug 23, 2013
Provisional Application 61694756 · Aug 29, 2012
Related Publication 20230384302A1 · Nov 30, 2023
References Cited (138)
US 3917538A · Rosensweig · 1975 [cited by applicant]
US 3976197A · Bhattacharya · 1976 [cited by applicant]
US 4083957A · Lang · 1978 [cited by applicant]
US 4092229A · Bhattacharya · 1978 [cited by applicant]
US 4276139A · Lawson · 1981 [cited by applicant]
US 4554088A · Whitehead et al. · 1985 [cited by applicant]
US 4698302A · Whitehead et al. · 1987 [cited by applicant]
US 4927749A · Dorn · 1990 [cited by applicant]
US 5466574A · Liberti et al. · 1995 [cited by applicant]
US 5466587A · Fitzpatrick-McElligott et al. · 1995 [cited by applicant]
US 5597531A · Liberti et al. · 1997 [cited by applicant]
US 6153373A · Benjamin et al. · 2000 [cited by applicant]
US 7169548B2 · Maxwell et al. · 2007 [cited by applicant]
US 7699979B2 · Li et al. · 2010 [cited by applicant]
US 7754444B2 · Xu et al. · 2010 [cited by applicant]
US 7776580B2 · Zhang et al. · 2010 [cited by applicant]
US 7829350B2 · Josephson et al. · 2010 [cited by applicant]
US 9804153B2 · Krug · 2017 [cited by examiner]
US 10324086B2 · Krug · 2019 [cited by examiner]
US 11531026B2 · Krug · 2022 [cited by examiner]
US 11630108B2 · Krug · 2023 [cited by examiner]
US 11662344B2 · Krug · 2023 [cited by examiner]
US 20020034537A1 · Schulze et al. · 2002 [cited by applicant]
US 20020182751A1 · Herr et al. · 2002 [cited by applicant]
US 20040142384A1 · Cohen et al. · 2004 [cited by applicant]
US 20050025971A1 · Cho et al. · 2005 [cited by applicant]
US 20060141512A1 · Sinha et al. · 2006 [cited by applicant]
US 20080044811A1 · Haugland · 2008 [cited by applicant]
US 20080206771A1 · Liu · 2008 [cited by applicant]
US 20080318250A1 · Gilmer et al. · 2008 [cited by applicant]
US 20090023205A1 · Dennig et al. · 2009 [cited by applicant]
US 20090042255A1 · Liu · 2009 [cited by applicant]
US 20090101507A1 · Aitken et al. · 2009 [cited by applicant]
US 20090208977A1 · Hudson et al. · 2009 [cited by applicant]
US 20090306461A1 · Oksenberg et al. · 2009 [cited by applicant]
US 20100081130A1 · Lee et al. · 2010 [cited by applicant]
US 20100129808A1 · Mirkin et al. · 2010 [cited by applicant]
US 20100200405A1 · Lenz · 2010 [cited by applicant]
US 20110086336A1 · Herickhoff et al. · 2011 [cited by applicant]
US 20110201047A1 · Paduch · 2011 [cited by applicant]
US 20110256581A1 · Gregory et al. · 2011 [cited by applicant]
US 20120100546A1 · Lowery, Jr. et al. · 2012 [cited by applicant]
US 20120270204A1 · Fox · 2012 [cited by examiner]
EP 0113452A2 · 1984 [cited by applicant]
FR 2811682A3 · 2002 [cited by applicant]
FR 2827799A1 · 2003 [cited by applicant]
JP 53056381A · 1978 [cited by applicant]
JP 601564A · 1985 [cited by applicant]
JP 2004073112A · 2004 [cited by applicant]
JP 2012037259A · 2012 [cited by applicant]
WO 8806632A1 · 1988 [cited by applicant]
WO 9007380A2 · 1990 [cited by applicant]
WO 9741224A1 · 1997 [cited by applicant]
WO 9958977A1 · 1999 [cited by applicant]
WO 2004029221A1 · 2004 [cited by applicant]
WO 2005054808A2 · 2005 [cited by applicant]
WO 2008095155A2 · 2008 [cited by applicant]
WO 2010073014A1 · 2010 [cited by applicant]
Chan et al. A simple zeta method for sperm selection based on membrane charge. Fertility and Sterility 85 (2): 481-486 (Feb. 2006). [cited by examiner]
International Search Report for PCT/US13/56526 (Feb. 7, 2014). [cited by applicant]
Written Opinion for PCT/US13/56526 (Feb. 7, 2014). [cited by applicant]
Shannon, P. “Contribution of Seminal Plasma, Sperm Numbers and Gas Phase to Dilution Effects of Bovine Spermatozoa.” J. Dairy Sci., 48: 1357 (1965). [cited by applicant]
Shannon, P. et al. “Toxic Effect and Action of Dead Sperm on Diluted Bovine Semen.” J. Dairy Sci., 55: 615-620 (1972). [cited by applicant]
Lindemann, C. B. et al. “A Comparative Study of the Effects of Freezing and Frozen Storage on Intact and Demembranated Bull Spermatozoa.” Cryobiology, 19: 20-28 (1982). [cited by applicant]
Saacke, R. G. et al. “Semen Quality Tests and Their Relationship to Fertility.” Proc. 4th National Association of Animal Breeders, Tech. Conf. Artificial Insemination and Reproduction, Apr. 18-20, 1972, Madison, WI, Nat… [cited by applicant]
G.W. Salisbury, G. W. et al. “Physiology of Reproduction and Artificial Insemination of Cattle.” second edition, Copyright 1961 & 1978. ISBN 0-7167-0025-5. [cited by applicant]
Schenk, J. L. et al. “Cryopreservation of Flow-Sorted Bovine Spermatozoa.”, Theriogenology 52:1375-1391, 1999. [cited by applicant]
Parish, J. J. et al. “Capacitation of bovine sperm by Heparin”, Biology of Reproduction 38:1171-1180, 1988. [cited by applicant]
Anzar, M. et al. “Filtration of Bovine Semen. I. Development of a Sephadex Ion-Exchange Filter.” Anim. Reprod. Sci. 1993; 31:187-195. [cited by applicant]
Avery, B. et al. “Impact of Percoll on Bovine Spermatozoa Used for in Vitro Insemination.” Theriogenology 1995; 44:871-788. [cited by applicant]
Correa, J. R. et al. “Preparation and Recovery of Frozen-Thawed Bovine Spermatozoa Via Various Sperm Selection Techniques Employed in Assisted Reproductive Technologies.” Theriogenology 1996; 46:1225-1232. [cited by applicant]
Garner, D. L.et al. “Dual DNA Staining Assessment of Bovine Sperm Viability Using SYBR-14 and Propidium Iodide.” J Andral 1994; 15:620-629. [cited by applicant]
Graham, E. F. et al. “The Effect of Whole Ejaculate Filtration on the Morphology and Fertility of Bovine Semen.” J. Dairy Sci. 1990; 73:91-97. [cited by applicant]
Johnson, L. A. et al. “Sex Preselection in Rabbits: Live births from X and Y Bearing Sperm Separated by DNA and Cell Sorting.” Biol Reprod 1989; 41:199-203. [cited by applicant]
Luderer, A. A. et al. “Separation of Bovine Spermatozoa by Density on Water Insoluble Newtonian Gels and Their Use or Insemination.” Biol Reprod 1982; 26 (5):813-824. [cited by applicant]
Pasteur, X. et al. “Identification of Two Human Sperm Populations Using Flow and Image Cytometer.” Molecular Reproduction and Development 1994; 38:303-309. [cited by applicant]
BcMagTm Carboxy-terminated Magnetic Beads. Product Manual. Bioclone, Inc. 2010 ittp://www.bioclone.us/files/BcMag_Carboxy-Terminated_Magnetic_Beads2.pdf. [cited by applicant]
BcMagTm Carboxy-terminated Magnetic Beads. Bioclone, Inc. 2010 http://www.bioclone.us/carboxyl-terminated-magnetic-beads-particle-resin-matrix.html. [cited by applicant]
BcMagTm SAX Magnetic Beads. Bioclone, Inc. 2010. http://www.bioclone.us/Strong-Anion-Exchange-magnetic-beads-particle-resin-matrix.html. [cited by applicant]
BcMagTm SAX Magnetic Beads. Product Manual. Bioclone, Inc. 2010 http://www.bioclone.us/files/BcMag_SAX_Magnetic_Beads.pdf. [cited by applicant]
Dynabeads SAX. Invitrogen Dynal Invitrogen Bead Separation; 2007. http://tools.thermofisher.com/content/sfs/manuals/105%2015D_16D_Dynabeads_SAX_rev002.pdf. [cited by applicant]
Dynabeads WCX. Invitrogen Dynal Invitrogen Bead Separation; 2008. http://tools.thermofisher.com/content/sfs/manuals/105%2011D12D%20%28001%29.pdf. [cited by applicant]
Dynabeads SCX. Invitrogen Dynal Invitrogen Bead Separation; 2007. http://tools.thermofishercom/content/sfs/manuals/105%2013D_14D%20Dynabeads%20SCX%28rev002%29.pdf. [cited by applicant]
M-Beads—Magnetic Silica Beads. MoBiTec; Magnetic Silica Beads. 2010. [cited by applicant]
Qiagen Purification Technologies. Qiagen anion-exchange, silica gel membrane, and magnetic particle technologies. 2012. [cited by applicant]
Propidium Iodide Staining Solution Technical Data Sheet. BD Pharmingen. BD Biosciences. 2006. [cited by applicant]
Dynabeads M-270 Carboxylic Acid. Invitrogen, 2012. https://tools.thermofishercom/content/sfs/manuals/dynabeads_m270carboxylicacid_man.pdf. [cited by applicant]
Australian Examination Report dated Oct. 27, 2015 for Appl. No. 2013309137. [cited by applicant]
Said et al. “Utility of Magnetic Cell Separation as a Molecular Sperm Preparation Technique.” J Andral. 2008 ; 29(2):134-142. [cited by applicant]
Hofmo, Peer Ola “Sperm Sorting and Low-Dose Insemination in the Pig—An Update” Acta Veterinaria Scandinavica 2006, 48(Suppl 1):S11. [cited by applicant]
Clemente, Henry. “Does Mammalian Sperm have a Charge.” ResearchGate. 2013. http://www.researchgate.net/post/Does_mammalian_sperm_have_a_charge. [cited by applicant]
Peter et al. “Fractionation of Bovine Spermatozoa for. Sex Selection: A Rapid Immunomagnetic Technique to Remove Spermatozoa That Contain the H—Y Antigen.” Theriogenology 40:1177-1185, 1993. [cited by applicant]
EP Extended Search Report dated Feb. 17, 2016 for Appl. No. 13832402.5. [cited by applicant]
Zhaogang “Highly Magnetizable Superparamagnetic Iron Oxide Nanoparticles Embedded Mesoporous Silica Spheres and Their Application for Efficient Recovery of DNA from Agarose Gel” Journal of Materials Chemistry, vol. 19, … [cited by applicant]
Bruce I J et al: “Synthesis, Characterisation and Application of Silicamagnetite Nanocomposites”, Journal of Magnetism and Magnetic Materials, vol. 284, Dec. 1, 2004, pp. 145-160. [cited by applicant]
CA Examination Report dated Jan. 21, 2016 for Appl. No. 2,883,328. [cited by applicant]
JP Examination Report dated Feb. 29, 2016 for Appl. No. 2015-529882. [cited by applicant]
Van Wienen, Marjet et al. “Single Layer Centrifugation with Androcoll-P Can be Scaled-Up to Process Larger Volumes of Boar Semen.” ISRN Veterinary Science. vol. 2011, Article ID 548385. pp. 8. [cited by applicant]
AU Examination Report dated Apr. 8, 2016 for Appl. No. 2013309137. [cited by applicant]
RU Examination Report dated May 10, 2016 for Appl. No. 2015111211. [cited by applicant]
AU Examination Report dated Jul. 8, 2016 for Appl. No. 2013309137. [cited by applicant]
RU Examination Report dated Oct. 4, 2016 for Appl. No. 2015111211. [cited by applicant]
JP Examination Report dated Oct. 4, 2016 for Appl. No. 2015-529882. [cited by applicant]
NZ Examination Report dated Nov. 17, 2016 for Appl. No. 705665. [cited by applicant]
Khajavi et al. Can Zeta sperm selection method, recover sperm with higher DNA integrity compare to density gradient centrifugation. Iranian Journal of Reproductive Medicine 7 (2): 73-77 (2009). [cited by applicant]
CA Examination Report dated Nov. 25, 2016 for Appl. No. 2,883,328. [cited by applicant]
EP Examination Report dated Feb. 2, 2017 for Appl. No. 13832402.5. [cited by applicant]
New Zealand Notice of Acceptance issued on Aug. 21, 2017 for NZ Appl. No. 705665. [cited by applicant]
European Intention to Grant Issued on Aug. 30, 2017 in related EP Appl. No. 13832402.5. [cited by applicant]
Australian Examination Report issued on Nov. 6, 2017 in related AU Appl. No. 2016247202. [cited by applicant]
Canadian Examination Report issued on Nov. 20, 2017 in related CA Appl. No. 2883328. [cited by applicant]
Notice of Allowance issued on Aug. 16, 2017 in related U.S. Appl. No. 13/974,139. [cited by applicant]
Office Action issued on Sep. 15, 2015 in related U.S. Appl. No. 13/974,139. [cited by applicant]
Final Office Action issued on Mar. 31, 2016 in related U.S. Appl. No. 13/974,139. [cited by applicant]
Office Action issued on Sep. 8, 2016 in related U.S. Appl. No. 13/974,139. [cited by applicant]
Final Office Action issued on Jan. 1, 2017 in related U.S. Appl. No. 13/974, 139. [cited by applicant]
Office Action issued on May 3, 2017 in related U.S. Appl. No. 13/974,139. [cited by applicant]
US Notice of Allowance issued on Aug. 16, 2017 for U.S. Appl. No. 13/974,139. [cited by applicant]
Australian Examination Report Issued on May 15, 2018 in related AU Appl. No. 2016247202. [cited by applicant]
US Office Action Issued on Jun. 8, 2018 in related U.S. Appl. No. 14/960,096. [cited by applicant]
Extended European Search Report Issued on Feb. 19, 2017 in related EP Appl. No. 17208185.3. [cited by applicant]
Office Action issued on Feb. 8, 2017 in related U.S. Appl. No. 14/960,096. [cited by applicant]
Said et al. “Effects of advanced selection methods on sperm quality and ART outcome: a systematic review.” Human Reproduction Update, vol. 17, No. 6, Nov. 2011, pp. 719-733. [cited by applicant]
European Decision to Grant Issued on Jan. 25, 2017 in related EP Appl. No. 13832402.5. [cited by applicant]
US Office Action Issued on Nov. 4, 2018 in related U.S. Appl. No. 14/960,096. [cited by applicant]
Indian Examination Report Issued on Dec. 6, 2018 in related IN Appl. No. 538/KOLNP/2015. [cited by applicant]
EPO Extended Search Report Issued on Dec. 12, 2018 in related EP Appl. No. 15866159.5. [cited by applicant]
Grunewald et al: “Enrichment of non-apoptotic human spermatozoa after cryopreservation by immunomagnetic cell sorting”, Cell and Tissue Banking, 2(3), pp. 127-133. [cited by applicant]
Glander et al: “Deterioration of spermatozoal plasma membrane is associated with an increase of sperm lyso-phosphatidylcholines”, Andrologia, 34(6), pp. 360-366. [cited by applicant]
Canadian Examination Reports issued on Apr. 14, 2021 in related CA Appl. No. 2,963,821. [cited by applicant]
Australian Examination report issued on May 28, 2020 in related AU Appl. No. 2015357516. [cited by applicant]
Australian Examination report issued on Jun. 10, 2020 in related AU Appl. No. 2019246835. [cited by applicant]
Brazil examination report issued on Jul. 9, 2020 in related BR Appl. No. BR112015004374-7. [cited by applicant]
European examination report issued on Jul. 6, 2020 in related EP Appl. No. 19180225.5. [cited by applicant]
US Office Action issued on Apr. 15, 2022 in related U.S. Appl. No. 16/438,215. [cited by applicant]
US Final Office Action issued on Aug. 16, 2022 in related U.S. Appl. No. 16/438,215. [cited by applicant]
US Notice of Allowance issued on Jan. 17, 2023 in related U.S. Appl. No. 16/438,215. [cited by applicant]
European Examination Report issued Aug. 29, 2024, in related EP Appl No. 22152556.1, filed on Jan. 20, 2022. [cited by applicant]