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In this case, the main objective is to modify the environment of the hapten to create an immunological response in vivo. A hapten is usually a small molecule that is not able to generate an immune response on its own, but can react with the products of such a response once generated. Most often these products are antibodies having binding specificity for the hapten. The complexities involved in achieving a successful conjugation strategy are best illustrated in the problems and concerns dealing with hapten–carrier conjugation.

0) that it is virtually always protonated and carries a positive charge. 1). Thus, at physiological pH, these residues contribute to the overall net positive charge of an intact protein molecule. The amine containing side chains in lysine, arginine, and histidine typically are exposed on the surface of proteins and can be derivatized with ease. 8). In alkylation, an active 10 1. 9 The mechanism of acylation proceeds through the attack of a nucleophile, generating a tetrahedral intermediate, which then goes on to form the product.

Most of these potential difficulties can be overcome through careful selection of the reaction conditions and through knowledge of the cofactor dependencies that are critical to the activity of the protein being modified. Post-translational modifications to protein structure are covalent changes that occur as the result of controlled enzymatic reactions or due to chemical reactions not under enzymatic regulation. One of the most common cellular modifications performed on proteins after ribosomal synthesis is glycosylation.