	US 12,285,469 B2
	Complex of botulinum neurotoxin E and its binding protein as a formulation with enhanced potency
Bal Ram Singh, Dartmouth, MA (US); and Raj Kumar, Dartmouth, MA (US)
Assigned to Institute of Advanced Sciences, Inc., North Dartmouth, MA (US)
Filed by Institute of Advanced Sciences, Inc., North Dartmouth, MA (US)
Filed on Mar. 11, 2024, as Appl. No. 18/601,211.
Application 18/601,211 is a division of application No. 17/200,822, filed on Mar. 13, 2021, granted, now 11,957,740.
Prior Publication US 2024/0216488 A1, Jul. 4, 2024
Int. Cl. A61K 38/48 (2006.01); C07K 1/14 (2006.01); C07K 1/18 (2006.01)

CPC A61K 38/4893 (2013.01) [C07K 1/145 (2013.01); C07K 1/18 (2013.01)]

10 Claims

1. A method for the isolation of an L complex or a M complex of a botulinum neurotoxin type E, comprising the steps of:

a) passing a solution comprising an L complex or an M complex of a botulinum neurotoxin type E through a functionalized ion exchange column comprising 2-(diethylamino)ethyl or acetate (O—CH₂COO); and

b) recovering the product;

wherein the M-complex comprises: a neurotoxin binding protein (NBP) bound to a Clostridium type E botulinum neurotoxin (BoNT/E), said M-complex with a molecular weight of about 250 kDa to about 300 kDa; and

wherein the L-complex comprises: the neurotoxin binding protein (NBP) bound to the Clostridium type E botulinum neurotoxin (BoNT/E), and a neurotoxin associated protein of 80, 60, 40, and 18 kDa (NAP-80; NAP-60, NAP-40, and NAP-18).