Systems and methods for testing and screening using compound bound substrates

A method of testing a sample for the presence of antibodies reactive to pathogen-inactivated red blood cells, the method comprising:

• a) providing a sample from a potential recipient of an infusion of the pathogen-inactivated red blood cells, wherein the sample comprises serum or plasma;
• b) contacting the sample with red blood cells, wherein a moiety is bound to the surface of the red blood cells;
• and wherein the moiety is selected from the group consisting of compounds of any of the following Formulae:
• wherein at least one of R1, R2, R3, R4, R5, R6, R7, R8 and R9 is —V—X-E,
• wherein V is independently —R11—, —NH—R11— or —N(CH3)—R11—, where —R11— is independently —C1-8alkyl-, —C1-8heteroalkyl-, -aryl-, -heteroaryl-, —C1-3alkyl-aryl-, —C1-3heteroalkyl -aryl-, —C1-3alkyl-heteroaryl-, —C1-3heteroalkyl-heteroaryl-, -aryl-C1-3alkyl-, -aryl-C1-3heteroalkyl-, -heteroaryl-C1-3alkyl-, -heteroaryl-C1-3heteroalkyl-, —C1-3alkyl-aryl-C1-3 alkyl-, —C1-3heteroalkyl-aryl-C1-3 alkyl-, —C1-3alkyl-heteroaryl-C1-3 alkyl-, —C1-3alkyl-aryl-C1-3 heteroalkyl-, —C1-3heteroalkyl-heteroaryl-C1-3 alkyl-, —C1-3heteroalkyl-aryl-C1-3 heteroalkyl-, —C1-3alkyl-heteroaryl-C1-3 heteroalkyl-, or —C1-3heteroalkyl-heteroaryl-C1-3 heteroalkyl-;
• X is independently —R11—;
• and
• E is independently selected from the group consisting of —N(R12)2, —N(R12)(R13), —S—R12, and
• where —R12 is —CH2CH2-G, where each G is
• independently —Cl, —Br, —I, —O—S(═O)2—CH3, —O—S(═O)2—CH2—C6H5, or —O—S(═O)2—C6H4—CH3;
• and
• where R13 is independently —C1-8 alkyl, —C1-8 heteroalkyl, -aryl, -heteroaryl, —C1-3alkyl-aryl, —C1-3heteroalkyl-aryl, —C1-3alkyl-heteroaryl, —C1-3heteroalkyl-heteroaryl, -aryl-C1-3alkyl, -aryl-C1-3heteroalkyl, -heteroaryl-C1-3alkyl, -heteroaryl-C1-3 heteroalkyl, —C1-3alkyl-aryl-C1-3 alkyl, —C1-3heteroalkyl-aryl-C1-3 alkyl, —C1-3alkyl-heteroaryl-C1-3 alkyl, —C1-3alkyl-aryl-C1-3 heteroalkyl, —C1-3heteroalkyl-heteroaryl-C1-3 alkyl, —C1-3heteroalkyl-aryl-C1-3 heteroalkyl, —C1-3alkyl-heteroaryl-C1-3 heteroalkyl, or —C1-3heteroalkyl-heteroaryl-C1-3 heteroalkyl;
• and the remainder of R1, R2, R3, R4, R5, R6, R7, and R8 are independently selected from the group consisting
• of —H, —R10, —O—R10, —NO2, —NH2, —NH—R10, —N(R10)2, —F, —Cl, —Br, —I, —C(═O)—R10, —C(═O)—O—R10, and —O—C(═O)—R10,
• where —R10 is independently
• H, —C1-8alkyl, —C1-8heteroalkyl, -aryl, -heteroaryl, —C1-3alkyl-aryl, —C1-3heteroalkyl-aryl, —C1-3alkyl -heteroaryl, —C1-3heteroalkyl-heteroaryl, -aryl-C1-3alkyl, -aryl-C1-3heteroalkyl, -heteroaryl-C1-3alkyl, -heteroaryl-C1-3heteroalkyl, —C1-3alkyl-aryl-C1-3 alkyl, —C1-3heteroalkyl-aryl-C1-3 alkyl, —C1-3alkyl-heteroaryl-C1-3 alkyl, —C1-3alkyl-aryl-C1-3 heteroalkyl, —C1-3heteroalkyl-heteroaryl-C1-3 alkyl, —C1-3heteroalkyl-aryl-C1-3 heteroalkyl, —C1-3alkyl-heteroaryl-C1-3 heteroalkyl, or —C1-3heteroalkyl-heteroaryl-C1-3 heteroalkyl;
• where R1, R2, R3, R4, R5, R6, R7, and R8 are independently selected from the group consisting of —H, —R10, —O—R10, —NO2, —NH2, —NH—R10, —N(R10)2, —F, —Cl, —Br, —I, —C(═O)—R10, —C(═O)—O—R10, and —O—C(═O)—R10, where —R10 is independently H, —C1-8 alkyl, —C1-8 heteroalkyl, -aryl, -heteroaryl, —C1-3alkyl-aryl, —C1-3heteroalkyl-aryl, —C1-3alkyl-heteroaryl, —C1-3heteroalkyl-heteroaryl, -aryl-C1-3alkyl, -aryl-C1-3heteroalkyl, -heteroaryl-C1-3alkyl, -heteroaryl-C1-3 heteroalkyl, —C1-3alkyl-aryl-C1-3 alkyl, —C1-3heteroalkyl-aryl-C1-3 alkyl, —C1-3alkyl-heteroaryl-C1-3 alkyl, —C1-3alkyl-aryl-C1-3 heteroalkyl, —C1-3heteroalkyl-heteroaryl-C1-3 alkyl, —C1-3heteroalkyl-aryl-C1-3 heteroalkyl, —C1-3alkyl-heteroaryl-C1-3 heteroalkyl, or —C1-3heteroalkyl-heteroaryl-C1-3 heteroalkyl;
• R20 is —H or —CH3;
• and
• R21 is —R11—X-E, where —R11— is independently —C1-8alkyl-, -C18heteroalkyl-, -aryl-, -heteroaryl-, —C1-3alkyl-aryl-, —C1-3heteroalkyl -aryl-, —C1-3alkyl-heteroaryl-, —C1-3heteroalkyl-heteroaryl-, -aryl-C1-3alkyl-, -aryl-C1-3heteroalkyl-, -heteroaryl-C1-3alkyl-, -heteroaryl-C1-3heteroalkyl-, —C1-3alkyl-aryl-C1-3 alkyl-, —C1-3heteroalkyl-aryl-C1-3 alkyl-, —C1-3alkyl-heteroaryl-C1-3 alkyl-, —C1-3alkyl-aryl-C1-3 heteroalkyl-, —C1-3heteroalkyl-heteroaryl-C1-3 alkyl-, —C1-3heteroalkyl-aryl-C1-3 heteroalkyl-, —C1-3alkyl-heteroaryl-C1-3 heteroalkyl-, or —C1-3heteroalkyl-heteroaryl-C1-3 heteroalkyl-;
• X is independently —R11—;
• and
• E is independently selected from the group consisting of —N(R12)2, —N(R12)(R13), —S—R12, and
• where —R12 is —CH2CH2-G,
• where each G is independently —Cl, —Br, —I, —O—S(═O)2—CH3, —O—S(═O)2—CH2—C6H5, or —O—S(═O)2—C6H4—CH3;
• and where R13 is independently —C1-8 alkyl, —C1-8 heteroalkyl, -aryl, -heteroaryl, —C1-3alkyl-aryl, —C1-3heteroalkyl-aryl, —C1-3alkyl-heteroaryl, —C1-3heteroalkyl-heteroaryl, -aryl-C1-3alkyl, -aryl-C1-3heteroalkyl, -heteroaryl-C1-3alkyl, -heteroaryl-C1-3 heteroalkyl, —C1-3alkyl-aryl-C1-3 alkyl, —C1-3heteroalkyl-aryl-C1-3 alkyl, —C1-3alkyl-heteroaryl-C1-3 alkyl, —C1-3alkyl-aryl-C1-3 heteroalkyl, —C1-3heteroalkyl-heteroaryl-C1-3 alkyl, —C1-3heteroalkyl-aryl-C1-3 heteroalkyl, —C1-3alkyl-heteroaryl-C1-3 heteroalkyl, or —C1-3heteroalkyl-heteroaryl-C1-3 heteroalkyl;
• and derivatives thereof, or a salt or stereoisomer of any of the foregoing;
• c) assaying an amount of binding between antibodies from the sample and the surface-bound moiety of the red blood cells, wherein binding between the antibodies and the surface-bound moiety indicates presence of antibodies reactive to the pathogen-inactivated red blood cells in the potential recipient.