Hapten to protein

Table 7 Conjugation of an amino-containing hapten to protein using the diazotization method Materials...

The above methods were utilized to conjugate a carboxyl group on a hapten to an amino residue on a protein. Obviously, the above reactions could be, and have been, utilized to conjugate an amino residue on a hapten to the carboxyl residues on proteins. However, there are additional methods which have proven useful for conjugating amine containing haptens to proteins. ... 

The linkage of haptens to proteins generally occurs at the most... 

Some reagents for the conjugation of haptens to proteins (modified after Parker, 1976)... 

GE Abraham, PK Grover. Covalent linkage of hormonal haptens to protein carriers for use in radioimmunoassay. In WD Odell, WH Daughaday, eds. Principle of Competitive Protein-Binding Assays. Philadelphia Lippincott, 1974, p 134. 

Activity Many F. induce a brown pigmentation in the skin on exposure to sunlight. They are photosensitizing, phototoxic, and can induce allergic dermatitis (Berloque s dermatitis), possibly with severe symptoms such as blister formation. The effect can also arise when, for example, scented toilet waters whose plant components still contain F. are applied to the skin. Use In photochemotherapy for vitiligo, psoriasis, atopic dermatitis, mycosis togoides. Photoactivated F. bind covalently as haptens to proteins as well as pyrimidines of DNA and thus have antimitotic effects. For synthesis, see Lit.. ... 

Table 3 Conjugation of a carboxyl-containing hapten to a protein...

Hapten density is important for both immunization and assay performance, and hence the extent of conjugation or hapten density should be confirmed by established methods. A characteristic ultraviolet (UV) or visible absorbance spectrum that distinguishes the hapten from the carrier protein or use of a radiolabeled hapten can be used to determine the degree of conjugation. If the hapten has a similar A. iax to the protein, the extent of incorporation can still be estimated when the concentration of the protein and the spectral characteristics of the hapten and protein are known. The difference in absorbance between the conjugate and the starting protein is proportional to... 

Cohen et al., 2007), and an improved method for coupling synthetic peptide haptens to carrier proteins (Lateef et al., 2007). 

Figure 19.16 A common way of conjugating sulfhydryl-containing haptens to carrier proteins is to activate the carrier with sulfo-SMCC to create an intermediate maleimide derivative. The maleimide groups then can be coupled to thiols to form thioether bonds.

The crosslinking scheme using this method can make use of the native e- and N-terminal amines on carrier proteins as the source of primary amine for the condensation reaction. Added to the conjugation reaction then is formaldehyde and the desired hapten to be coupled containing an appropriately active hydrogen. 

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