| 摘要 |
This invention discloses “Artificially Cleaved Epitope (ACE)” methods, antibodies, reagents, immunoassays, and kits for designing and detecting hidden epitopes/antigens. The ACE methods can detect epitopes that are either absent or poorly accessible naturally to antibodies, and thus must be specifically and artificially created (free terminals) and/or exposed in samples and sample preparations for antibody detection. The ACE structures include, but are not limited to, macromolecule-to-macromolecule conjugation sites, and any types of linear hidden epitopes. The ACE methods comprise ACE antigen design and ACE antigen detection. The ACE methods, antibodies, reagents, immunoassays, and kits are useful in research and discovery, diagnostic, and therapeutic applications. In another aspect, the ACE methods can artificially and specifically expose hidden antigens while reducing the antibody non-specific bindings in all antibody-based applications. |
| 主权项 |
1. A method of detecting a hydrolysis-created Artificially Cleaved Epitope (ACE) structure in a sample, wherein the ACE structure is hidden in its intact or natural form in a polymeric molecule and is poorly accessible to antibodies, comprising steps of:
(i) designing an ACE structure of formula L1-L2---Ln, wherein L1 and Ln are each terminal residues that are artificially created via chemical bond-specific cleavage by a hydrolytic enzyme or hydrolytic agent, wherein the antibody specifically recognizes at least one of the L1 or Ln terminal residues or residues between L1 and Ln; (ii) synthesizing the ACE structure; (iii) making an antibody against the ACE structure; (iv) creating the ACE structure in the sample by treating the sample preparation with the hydrolytic enzyme or hydrolytic agent thereby exposing the formerly hidden ACE. structure to specific interaction with the antibody; and (v) detecting the ACE structure created in step (iv) with the antibody, wherein the antibody specifically binds to at least one of the artificially created L1 or Ln terminal residues or residue(s) between L1 and Ln. |
