Solid-phase peptide synthesis on proteins.
A new method for solid-phase peptide synthesis in which a protein is used as the solid support has been developed. Two aspects of the method have been demonstrated. The peptides H-Phe-Leu-Glu-Glu-Val-OH (1) and H-Leu-Leu-Ala-Gly-Val-OH (2), respectively, were synthesized on the amino groups of BSA via a cleavable linker, using the Fmoc group protecting scheme. The free peptides were obtained by cleavage from the protein with 95% TFA, precipitation in diethyl ether and additional work-up by either dialysis or centrifugation through a membrane followed by gel filtration. The identity of the products was determined by amino acid analysis and HPLC. The peptide-protein conjugates, H-Ser-Met-Asp-Thr-Ser-Asn-Lys-Glu-Glu-Lys-BSA (3) and H-Thr-Val-Leu-BTG (4), were obtained in the same manner, omitting the cleavable linker group. It was found that 35-50 peptide chains were conjugated per molecule BSA and BTG, respectively. Immunization of rabbits with conjugate 3 gave rise to peptide specific antibodies. This method will be useful for generation of sequence specific antibodies, since the peptide is conjugated to the carrier protein exclusively via its C-terminus, and will allow synthesis of highly specific peptide-protein conjugates.
Hansen
P R
PR
Center for Medical Biotechnology, Chemistry Department, Royal Veterinary and Agricultural University, Copenhagen, Denmark.
Holm
A
A
Houen
G
G
D000595:Amino Acid Sequence; D005069:Evaluation Studies as Topic; D008722:Methods; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012995:Solubility; D013499:Surface Properties; D013954:Thyroglobulin / Q000737:chemistry
D009842:Oligopeptides; D011506:Proteins; D012710:Serum Albumin, Bovine; D013954:Thyroglobulin