Cell detection with conjugates having an enzymatically releasable detection moiety
View Patent ↗The invention is directed to a method for detecting a target moiety in a sample of biological specimens by: a) providing at least one conjugate with the general formula (I) X n —P—Y m , with X is an detection moiety, P an enzymatically degradable spacer and Y an antigen recognizing moiety and n, m are integers between 1 and 100 and wherein X and Y are covalently bound to P; b) contacting the sample of biological specimens with at least one conjugate, thereby labeling the target moiety recognized by the antigen recognizing moiety Y; c) detecting the target moiety labeled with the conjugate with the detecting moiety X; and d) enzymatically degrading spacer P, thereby cleaving the detection moiety X from the conjugate. The method is useful to identify target moieties on the biological specimens. The biological specimens detected by the conjugate can be subsequently removed from the sample.
1. A method for detecting a target moiety in a sample of biological specimens by:
a) providing at least one conjugate with the general formula (I) X n —P—Y m , wherein X is a detection moiety, P is an enzymatically degradable spacer, Y is an antigen recognizing moiety and n, m are integers between 1 and 100 and wherein X and Y are covalently bound to P;
b) contacting the sample of the biological specimens with the at least one conjugate, thereby labelling the target moiety recognized by the antigen recognizing moiety Y;
c) detecting the target moiety labelled with the conjugate; and
d) enzymatically degrading the spacer P, thereby cleaving the detection moiety X from the conjugate,
wherein the detection moiety X is a fluorescent moiety, the enzymatically degradable spacer P is dextran, and the antigen recognizing moiety Y is Fab, and
wherein by enzymatically degrading the spacer P, the antigen recognizing moiety Y is also cleaved from the target moiety.
2. The method according to claim 1 , wherein the biological specimens are separated from the sample by at least one of electrostatic forces, piezoelectric forces and mechanical separation.