The generation of antibodies to the coupling agent used for peptide conjugation

Christophe Boeckler
Journal of Immunological Methods
This paper exame the possible experimental pitfalls due to the generation of antiobdies to the coupling agent used for peptide conjugation. Although it has been known for many years that antibodies can be generated by coupling agent (or cross linker) used to conjugate peptides to a carrier protein. In general these linkers are highly immunogenic. In a comparative study of four bifunctional reagents, Peeters et al. (1989) found that antisera to the peptide conjugates could show antibody titre as high as 104-105 against reagents such as MBS or SMCC. The spacer originating wtih SPDP induced very little specific antiobdy. In the same series of experiments, MHS which is more stable than SPDP, whoed a moderate immunogenicity as compared with MBS and SMCC. Glutaraldehjyde, EDC, MCS, MBS, bisdiazobenzidine were all found to induce a strong antibody response (Bernatowicz and Matsueda, 1986; Boersma et al., 1988; Briand et al., 1985; Deen et al., 1990; Lloyd Jones et al., 1989). It is therefore advisable to prepare two conjugates of the same peptide with different carrier proteins and different coupling methods which will be respectively used for immunization and screeing of antibodies.

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