Simultaneous multiple synthesis of peptide-carrier conjugates.
Recently, the multipin approach for simultaneous multiple peptide synthesis was applied to the analysis of T cell determinants by using a novel cleavage method (Maeji et al., 1990). A diketopiperazine forming linker allowed cleavage of peptides into aqueous buffer which, without further purification, could be used immediately in cell culture assays. Another potential application of the technique is the simultaneous cleavage and coupling of peptides to immunogenic carriers. Without further purification the resulting conjugates can be used for the production of antipeptide antisera. The choice of carrier and conjugation chemistry is not restricted as peptide/pin cleavage occurs in aqueous solution over a range of pH and ionic strength. The method was assessed using the 2,4-dinitrophenyl group as a model hapten, diphtheria toxoid as the carrier, and N-(epsilon-maleimidocaproyloxy)succinimide as the cross-linking reagent. The resulting DNP-DT conjugate was used to prepare high titered specific anti-DNP antisera in mice.
Maeji
N J
NJ
Chiron Mimotopes Pty. Ltd., Clayton, Victoria, Australia.
Tribbick
G
G
Bray
A M
AM
Geysen
H M
HM
D019297:2,4-Dinitrophenol; D000818:Animals; D003432:Cross-Linking Reagents; D004140:Dinitrophenols / Q000276:immunology*; D004168:Diphtheria Toxoid; D006241:Haptens / Q000096:biosynthesis; D006863:Hydrogen-Ion Concentration; D007114:Immunization; D066298:In Vitro Techniques; D051379:Mice; D010455:Peptides / Q000138:chemical synthesis*; D013388:Succinimides / Q000276:immunology*; D015091:beta-Alanine / Q000032:analysis
D003432:Cross-Linking Reagents; D004140:Dinitrophenols; D004168:Diphtheria Toxoid; D006241:Haptens; D010455:Peptides; D013388:Succinimides; D015091:beta-Alanine; C072682:N-(epsilon-maleimidocaproyloxy)succinimide; D019297:2,4-Dinitrophenol