US 7,320,878 B2 | ||
Protease assay for therapeutic drug monitoring | ||
Sergei Gulnik, Frederick, Md. (US); Betty Yu, Frederick, Md. (US); John W Erickson, Potomac, Md. (US); and Martin Markowitz, New York, N.Y. (US) | ||
Assigned to Tibotec Pharmaceuticals, Ltd., (Iran, Islamic Republic of); Aaron Diamond Aids Research Center, New York, N.Y. (US); and The Rockefeller University, New York, N.Y. (US) | ||
Appl. No. 10/494,903 PCT Filed Nov. 08, 2002, PCT No. PCT/EP02/12631 § 371(c)(1), (2), (4) Date Nov. 04, 2004, PCT Pub. No. WO03/040390, PCT Pub. Date May 15, 2003. |
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Claims priority of provisional application 60/331117, filed on Nov. 08, 2001. | ||
Prior Publication US 2005/0221286 A1, Oct. 06, 2005 | ||
Int. Cl. C12Q 1/37 (2006.01); G01N 33/53 (2006.01) |
U.S. Cl. 435—23 [435/24; 435/7.72] | 16 Claims |
1. A method for determining the inhibitory potency of an HIV inhibitor in a biological sample, comprising:
i) providing a biological sample comprising said HIV inhibitor;
ii) providing HIV protease;
iii) providing a substrate for HIV protease R-E(EDANS)-S-Q-N-Y-P-I-V-Q-K(DABCY-L)-R-OH (SEQ ID NO: 10);
iv) separating HIV inhibitor bound to proteins in the biological sample from unbound HIV inhibitor by gel filtration, filters,
filter papers, special microtiter plates, dialysis, heating, precipitation, centrifugation, or antibodies or by combinations
thereof;
v) adding the resulting biological sample from which bound HIV inhibitor has been separated, the HIV protease and the HIV
protease substrate to a container;
vi) determining a signal; and
vii) relating the signal of vi) to a reference standard curve prepared with at least one reference.
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