	US 11,874,280 B2
	Use of divalent metals for enhancement of fluorescent signals
Melissa Jackson, New York, NY (US); Christopher Boyce, New York, NY (US); Eriko Matsui, New York, NY (US); Michael Vanbrunt, Covington, WA (US); Sharat Singh, Rancho Santa Fe, CA (US); and Tracy Matray, Snohomish, WA (US)
Assigned to Sony Group Corporation, Tokyo (JP)
Appl. No. 16/982,341
Filed by SONY GROUP CORPORATION, Tokyo (JP)
PCT Filed Mar. 8, 2019, PCT No. PCT/US2019/021323 § 371(c)(1), (2) Date Sep. 18, 2020, PCT Pub. No. WO2019/182765, PCT Pub. Date Sep. 26, 2019.
Claims priority of provisional application 62/645,121, filed on Mar. 19, 2018.
Prior Publication US 2021/0109104 A1, Apr. 15, 2021
Int. Cl. G01N 33/58 (2006.01); G01N 33/533 (2006.01)

CPC G01N 33/582 (2013.01) [G01N 33/533 (2013.01)]

23 Claims

1. A method for forming a covalent conjugate of a polymeric dye and a targeting moiety, the method comprising preparing a mixture comprising the polymeric dye, the targeting moiety, and a magnesium salt, and allowing the mixture to age for a time and at a temperature sufficient to form the covalent conjugate, wherein:

a) the polymeric dye comprises:

i) two or more fluorescent or colored moieties;

ii) at least one negatively charged phosphate group; and

iii) a reactive group Q capable of forming a covalent bond with a complementary reactive group Q′ on the targeting moiety,

wherein each of the two or more fluorescent or colored moieties are joined to an adjacent fluorescent or colored moiety via a linker comprising the at least one negatively charged phosphate group; and

b) the targeting moiety has affinity for a target analyte and comprises the complementary reactive group Q′.