Active-Hydrogen-Mediated Hapten—Carrier Conjugation

Active-Hydrogen-Mediated Hapten-Carrier Conjugation 

Conjugation chemistry for the coupling of haptens to carrier molecules is fairly well defined for compounds having common functional groups to facilitate such attachment. The types of functional groups generally useful for this operation include easily reactive components such as primary amines, carboxylic acids, aldehydes, or sulf-hydryls. 

Derivatives of carbohydrate antigens also have been coupled to carrier proteins 

Diazo linkages are reversible bonds that may be cleaved by addition of 0.1 M 

Add 20 ul of Ellman s reagent (lmg/ml dissolved in the gel filtration purification buffer) to each well including one containing only buffer (220 pi) to use as a blank. 

Measure the absorbance of all wells using a microplate reader with a filter set at 410nm. 

Diazo Cross-linked Hapten-Carrier Conjugate 

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Active-Hydrogen-Mediated Hapten-Carrier Conjugation... 

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