Zwitterionic reagents for labeling, cross-linking and improving the performance of chemiluminescent immunoassays.
Summary of "Zwitterionic reagents for labeling, cross-linking and improving the performance of chemiluminescent immunoassays."
Improving reagent performance in immunoassays both to enhance assay sensitivity and to minimize interference are ongoing challenges in clinical diagnostics. We describe herein the syntheses of a new class of hydrophilic reagents containing sulfobetaine zwitterions and their applications. These zwitterionic reagents are potentially useful for improving the properties of immunoassay reagents. We demonstrate for the first time that zwitterion labeling is a general and viable strategy for reducing the non-specific binding of proteins to microparticles and, to improve the aqueous solubility of hydrophobic peptides. We also describe the synthesis of zwitterionic cross-linking reagents and demonstrate their utility for peptide conjugation. In automated, chemiluminescent immunoassays, improved assay performance was observed for a hydrophobic, small analyte (theophylline) using an acridinium ester conjugate with a zwitterionic sulfobetaine linker compared to a hexa(ethylene)glycol linker. Sandwich assay performance for a large analyte (thyroid stimulating hormone) was similar for the two acridinium ester labels. These results indicate that zwitterions are complementary to poly(ethylene)glycol in improving the aqueous solubility and reducing the non-specific binding of chemiluminescent acridinium ester conjugates.
Affiliation
Siemens Healthcare Diagnostics, Advanced Technology and Pre-Development, 333 Coney Street, East Walpole, MA 02032, USA. anand.natrajan@siemens.com.
Journal Details
This article was published in the following journal.
Name: Organic & biomolecular chemistry
ISSN: 1477-0539
Pages: 1883-95
Links
- PubMed Source: http://www.ncbi.nlm.nih.gov/pubmed/22278720
- DOI: http://dx.doi.org/10.1039/c2ob06807a
Medical and Biotech [MESH] Definitions
Cross-linking Reagents
Reagents with two reactive groups, usually at opposite ends of the molecule, that are capable of reacting with and thereby forming bridges between side chains of amino acids in proteins; the locations of naturally reactive areas within proteins can thereby be identified; may also be used for other macromolecules, like glycoproteins, nucleic acids, or other.
Glutaral
One of the protein CROSS-LINKING REAGENTS that is used as a disinfectant for sterilization of heat-sensitive equipment and as a laboratory reagent, especially as a fixative.
Dinitrofluorobenzene
Irritants and reagents for labeling terminal amino acid groups.
Muramoylpentapeptide Carboxypeptidase
Enzyme which catalyzes the peptide cross-linking of nascent CELL WALL; PEPTIDOGLYCAN.
Mitomycin
An antineoplastic antibiotic produced by Streptomyces caespitosus. It is one of the bi- or tri-functional ALKYLATING AGENTS causing cross-linking of DNA and inhibition of DNA synthesis.
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