Antibody pair screening methods
View Patent ↗The invention provides methods for identifying antibody preparations that can form a pair of antibodies that optimally detect a target antigen, for example, in a sandwich immunoassay. These methods provide high affinity and epitope-specific antibodies.
1. A kit for performing an immunoassay to detect a C-reactive protein antigen in a test sample, the kit comprising a first monoclonal antibody and a second monoclonal antibody immobilized on a surface, the first antibody and the second antibody having a high specific affinity for the C-reactive protein antigen such that the first antibody and the second antibody form a ternary sandwich with the C-reactive protein antigen, wherein the first antibody and the second antibody do not exhibit substantial cross-reactivity with one another; wherein the percent specific activity of the ternary sandwich is from about 70% to about 99%, wherein the percent specific activity of the ternary sandwich is defined by the formula:
100×[(A−C)/A]
wherein
A is the ternary affinity value of the ternary sandwich; and
C is the cross-reactivity value between the first antibody and the second antibody.
2. The kit of claim 1 , the kit further comprising a container containing the antigen.
3. The kit of claim 1 , wherein the percent specific activity of the ternary sandwich is from about 90% to about 99%.
4. The kit of claim 1 , wherein the cross-reactivity value is less than about 15 SPR (surface plasmon resonance) resonance units.
5. The kit of claim 1 , wherein the cross-reactivity value is less than 15.4 SPR resonance units.
6. The kit of claim 1 , wherein the ternary affinity value is greater than about 460 SPR resonance units.
7. The kit of claim 1 , wherein the ternary affinity value is between about 460 SPR resonance units and about 1240 SPR resonance units.
8. The kit of claim 1 , wherein the ternary affinity value is between 460.7 SPR resonance units and 1237.3 SPR resonance units.