A micropreparation of fluorescein conjugates of immunoglobulin G and fab from serum

Joy G. Mohanty, Ken S. Rosenthal

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Protein A-Sepharose CL-4B (PAS) was used to isolate rabbit immunoglobulin G from crude anti-herpes simplex virus-1 serum. Papain treatment of the PAS-bound immunoglobulin G released Fab fragments from the solid support, while Fc-containing fragments remained bound to PAS. PAS-immobilized immunoglobulin G was fluoresceinated by reaction with fluorescein isothiocyanate followed by papain cleavage to yield fluorescein-conjugated Fab fragments in solution. These fragments retained activity toward herpes simplex virus-1 infected Vero cells as evaluated by immunofluorescence. This novel procedure represents the fastest and simplest method for preparing Fab or fluoresceinated Fab fragments directly from any volume of immune serum.

Original languageEnglish (US)
Pages (from-to)361-365
Number of pages5
JournalAnalytical Biochemistry
Volume146
Issue number2
DOIs
StatePublished - May 1 1985
Externally publishedYes

Keywords

  • Fab
  • HSV
  • IgG
  • fluorescein-IgG
  • protein A-Sepharose

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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