US 11,740,233 B2
Antibody-nanoparticle conjugates and methods for making and using such conjugates
Julia Ashworth-Sharpe, Tucson, AZ (US); Christopher Bieniarz, Tucson, AZ (US); Michael Farrell, Tucson, AZ (US); Donald D. Johnson, Tucson, AZ (US); Jerome W. Kosmeder, Tucson, AZ (US); Adrian E. Murillo, Tucson, AZ (US); Chol Steven Yun, San Diego, CA (US); and Zhanna Zhilina, Tucson, AZ (US)
Assigned to Ventana Medical Systems, Inc., Tucson, AZ (US)
Filed by Ventana Medical Systems, Inc., Tucson, AZ (US)
Filed on Jun. 21, 2018, as Appl. No. 16/14,965.
Application 14/691,826 is a division of application No. 13/640,944, granted, now 9,040,310, issued on May 26, 2015, previously published as PCT/US2011/034190, filed on Apr. 27, 2011.
Application 16/014,965 is a continuation of application No. 15/263,221, filed on Sep. 12, 2016, granted, now 10,031,134.
Application 15/263,221 is a continuation of application No. 14/691,826, filed on Apr. 21, 2015, granted, now 9,442,107, issued on Sep. 13, 2016.
Claims priority of provisional application 61/328,494, filed on Apr. 27, 2010.
Prior Publication US 2018/0372733 A1, Dec. 27, 2018
Int. Cl. G01N 33/543 (2006.01); G01N 33/531 (2006.01); G01N 33/68 (2006.01); G01N 33/553 (2006.01); C07F 9/12 (2006.01); C07F 9/50 (2006.01); C07F 9/572 (2006.01); C07F 9/655 (2006.01); G01N 33/58 (2006.01)
CPC G01N 33/54346 (2013.01) [C07F 9/12 (2013.01); C07F 9/5004 (2013.01); C07F 9/5022 (2013.01); C07F 9/5045 (2013.01); C07F 9/5728 (2013.01); C07F 9/65515 (2013.01); C07F 9/65517 (2013.01); C07F 9/65522 (2013.01); G01N 33/531 (2013.01); G01N 33/553 (2013.01); G01N 33/587 (2013.01); G01N 33/6854 (2013.01); G01N 2333/916 (2013.01)] 19 Claims
 
1. A method for producing an antibody-nanoparticle conjugate, comprising:
Providing a reduced intact antibody comprising 2 heavy chains and 2 light chains and having at least one thiol group, wherein the 2 heavy chains are covalently coupled to each other through at least one interchain disulfide bond; the reduced intact antibody is produced by reducing an intact antibody comprising two heavy chains and two light chains with dithiothreitol (DTT) under condition such that at least one thiol group is formed by reduction of at least one of a native interchain disulfide or a native intrachain disulfide of the intact antibody wherein the 2 heavy chains of the reduced intact antibody are covalently coupled to each other through at least one interchain disulfide bond; and
reacting the reduced intact antibody with an arylphosphine-nanoparticle composite and coupling the reduced intact antibody directly without an intervening linker to the arylphosphine-nanoparticle composite through the at least one formed thiol group through a metal-thiol bond, thereby producing the antibody-nanoparticle conjugate.