A synthetic strategy for simultaneous purification-conjugation of antigenic peptides.
A novel approach to the preparation of immunopeptide-carrier protein conjugates of improved chemical definition based on a solid-phase synthetic protocol combining incorporation of a Cys(Npys) N-terminal residue with systematic acetylation after every coupling, is described. The potential of the method is demonstrated in a synthesis of the tridecapeptide (Npys)-Cys-Val-Asn-Tyr-Ile-Arg-Lys-Arg-Ser-Leu-Gln-Thr-Val-OH in which the main product is purposely contaminated by a number of shorter truncated sequences resulting from intentionally defective couplings. From this peptide crude, and rather independently of its complexity, the target sequence can be selectively recovered and attached to a carrier molecule through a disulfide bond formed by reaction of the Npys-protected cysteine residue and a thiol function in the carrier. The process can be properly named purification-by-conjugation.
Ponsati
B
B
Department of Organic Chemistry, University of Barcelona, Spain.
Giralt
E
E
Andreu
D
D
D000595:Amino Acid Sequence; D000941:Antigens / Q000032:analysis*; D001894:Borohydrides; D002352:Carrier Proteins / Q000032:analysis / Q000138:chemical synthesis; D002851:Chromatography, High Pressure Liquid; D006433:Hemocyanins; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis / Q000138:chemical synthesis / Q000302:isolation & purification; D010455:Peptides / Q000138:chemical synthesis* / Q000276:immunology / Q000302:isolation & purification; D011485:Protein Binding; D012685:Sepharose; D012710:Serum Albumin, Bovine / Q000032:analysis
D000941:Antigens; D001894:Borohydrides; D002352:Carrier Proteins; D010446:Peptide Fragments; D010455:Peptides; D012710:Serum Albumin, Bovine; C025364:sodium borohydride; D012685:Sepharose; D006433:Hemocyanins