Use of commercial anion-exchange resins as solid support for peptide synthesis and affinity chromatography.
This report demonstrates that due to the presence of residual reactive sites in their matrices, classical diethylaminoethyl-attaching commercial anion-exchanger resins such as DEAE-MacroPrep and DEAE-Sephadex A50 supports can be used for peptide synthesis. Moreover, due to the high stability of the peptide-resin bond in the final cleavage treatments, desired peptidyl-resins free of side-chain protecting groups, which enables them to be further used as solid support for affinity chromatography, can be obtained. To demonstrate this potentiality, a fragment corresponding to the antigenic and immunodominant epitope of sporozoites of the Plasmodium falciparum malaria parasite was synthesized in these traditional resins and antibody molecules generated against the peptide sequence were successfully retained in these peptidyl supports. Due to the maintenance of their original anion-exchange capacities, the present findings open the unique possibility of applying, simultaneously, dual anion-exchange and affinity procedures for purification of a variety of macromolecules.
Nakaie
Clovis R
CR
Department of Biophysics, Universidade Federal de São Paulo, Rua 3 de Maio 100, CEP 04044-020 São Paulo, SP, Brazil. clovis.biof@epm.br
Ianzer
Danielle A
DA
Malavolta
Luciana
L
Cilli
Eduardo M
EM
Rodrigues
Mauricio M
MM
D000804:Angiotensin II / Q000138:chemical synthesis; D000837:Anion Exchange Resins / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification; D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D003637:DEAE-Dextran / Q000031:analogs & derivatives* / Q000737:chemistry; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D010452:Peptide Biosynthesis; D010446:Peptide Fragments / Q000138:chemical synthesis; D012117:Resins, Synthetic / Q000737:chemistry
D000837:Anion Exchange Resins; D000911:Antibodies, Monoclonal; D009842:Oligopeptides; D010446:Peptide Fragments; D012117:Resins, Synthetic; D000804:Angiotensin II; C027184:DEAE-Sephadex A-50; D003637:DEAE-Dextran; C052953:(asparaginyl-alanyl-asparaginyl-proline)3