Second-generation hapten 3

Development of such idiotype-dependent assays requires much labor and a long period because two-step antibody production is necessary first, immunization with a hapten-carrier conjugate to produce a specific anti-hapten antibody, then a second immunization with the anti-hapten antibody for generating two types of anti-idiotype antibodies. Furthermore, a suitable combination between a-type and /3-type antibodies, whose binding to the anti-hapten antibody is competitive, has to be found. 

Standard immunizations of mouse involve two injections, or boosts , of a hapten-carrier protein conjugate together with an adjuvant at a 15 days interval. At least eight weeks after the second boost, immune cells are stimulated by a final intravenous injection of hapten-conjugate, and the fusion carried out 4 days later to generate hybridoma from spleen cells (Fig. 7). Experiments with catalytic polyclonal antibodies [103] have shown that over-immunization re-... 

Amino-l-cyclohexylmethylphosphonic acid diethyl ester was utilized for the preparation of hapten 1, recently employed to induce the first monoclonal catalytic antibodies capable of catalyzing peptide bond formation[97]. The key steps of the synthesis of 1 include (a) preparation of scalemic a-aminophosphonate, (b) generation of the p-nitrobenzyl ester, (c) installation of a suitable second phosphonate ester linkage, (d) M-acylation with glutaric anhydride. 

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