Protein antigens conjugation with haptens

Dissolve the antigen (pure protein or hapten-carrier conjugate) with a concentration between 0.1 and 50 pg/ml in Soln. A. Pipet 0.1-0.2 ml of the antigen solution into the wells of a 96-well microtiter plate (high binding capacity) and incubate on a shaker at RT for 1 h or at 4 °C in a humid chamber overnight. 

Figure 7.80 The reaction of protein with the penicillin molecule (hapten) to form a possibly antigenic conjugate. Breakdown products may also react with protein. The protein might be either soluble or cellular. For penicillin G, R=benzyl. The strained p-lactam ring is indicated.

The nature of the specificity of antihapten antibodies can be demonstrated with a few examples of experimental results. Tables 2.1 and 2.2 present data on the specificity of rabbit antibody directed to the p,p -azophenylazobenzoate hapten group (2a). The immunizing antigen consisted of bovine serum proteins to which several hapten groups, per molecule of protein, had been conjugated by diazotization. (See Fig. 2.3.)... 

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