US 7,611,860 B2
Indicator for in-situ detecting of lysozyme
Drew M Ferguson, Cambridge (United Kingdom); Guy D. Milan, Great Shelford (United Kingdom); Crawford S. Dow, Coventry (United Kingdom); and Uthaya Swoboda, Coventry (United Kingdom)
Assigned to Cambridge Meditech Limited, Cambridge (United Kingdom)
Appl. No. 10/381,426
PCT Filed Oct. 15, 2001, PCT No. PCT/GB01/04588
§ 371(c)(1), (2), (4) Date Jul. 15, 2003,
PCT Pub. No. WO02/30478, PCT Pub. Date Apr. 18, 2002.
Prior Publication US 2004/0043422 A1, Mar. 04, 2004
Int. Cl. C08L 5/08 (2006.01); G01N 33/53 (2006.01); C12Q 1/34 (2006.01)
U.S. Cl. 435—18  [435/7.2; 106/162.1] 28 Claims
 
1. An indicator suitable for detecting at a location the presence of lysozyme or the presence of nicotinamide adenine dinucleotide (NADH) in the presence of lysozyme, wherein the indicator comprises:
(a) a biopolymer layer which is susceptible to degradation by lysozyme; and
(b) a signaling layer which comprises an indicator substance, which layer is adapted to produce a detectable signal upon contact with lysozyme or upon contact with NADH in the presence of lysozyme;
wherein, in use, the signaling layer (b) is at least initially protected from contact with NADH by the layer (a), and wherein
the signaling layer (b) is adapted to produce a detectable signal whose initial strength is proportional to the amount of NADH or lysozyme, and wherein the signaling layer (b) further comprises a pH indicator or assay reagents for detecting an additional enzyme, an immune cell or a micro-organism.