Recombinant peptides as immunogens: a comparison of protocols for antisera production using the pGEX system.
Biotechniques
; 12(4): 544-9, 1992 Apr.
Article
em En
| MEDLINE
| ID: mdl-1503757
ABSTRACT
Using an inducible vector system that directs high-level production and rapid purification of recombinant protein, we have immunized mice with peptides prepared by several methods:
1) samples fractionated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) subsequently transferred to PVDF membrane and subcutaneously implanted in mice; 2) samples cut directly from SDS-PAGE gels and injected intraperitoneally; 3) injection of recombinant protein bound to agarose beads; and 4) injection of log-phase E. coli transformed with recombinant vector. All four strategies yielded specific antisera reacting with both the parental fusion protein and the recombinant fragment as determined by enzyme-linked immunosorbent assay and immunoblot analysis. Specific recognition of the recombinant fragment was demonstrated by a competitive inhibition assay in which the parental fusion protein abrogated reactivity of serum with the isolated recombinant fragment.
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Base de dados:
MEDLINE
Assunto principal:
Vacinas Sintéticas
/
Imunização
/
Soros Imunes
Limite:
Animals
Idioma:
En
Revista:
Biotechniques
Ano de publicação:
1992
Tipo de documento:
Article