Submitted to: International Conference on Agri-Food Antibodies
Publication Type: Abstract Only
Publication Acceptance Date: August 26, 2005
Publication Date: September 6, 2005
Citation: Brandon, D.L., Binder, R.G. 2005. 2-morpholinoethylisocyanide as a coupling agent for hapten-protein conjugates [abstract]. International Conference on Agri-Food Antibodies, 6-9 September 2005, Chester, UK Abstract No.P-07. Technical Abstract: The conjugation of low molecular weight haptens to proteins and enzymes are essential steps in the production of antibodies and development of immunoassays for these compounds. The utility of water-soluble carbodiimides is generally limited by the low solubilities of some haptens in water and by the formation of N-acylurea and other by-products. These urea substituents may reduce the solubility of the conjugate and could act as unwanted haptens. Desirable protein coupling reactions, which can readily be done on a micro scale, involve facile purification procedures and conditions that do not denature proteins and enzymes. To address these problems, we used 2-morpholinoethylisocyanide to couple several carboxylic acid haptens to proteins and enzymes, including derivatives of cholesterol, anthelmintic benzimidazoles, and vitamins. The procedure is simple, involves only small volumes of water-miscible organic reagent, and produces conjugates that can be purified by dialysis.