Purification and Analysis of Immune Complexes with the Aid of Rheumatoid Factor-Coated Tubes

Z. Gilead, Y. Gazitt, G. Klein, D. Sulitzeanu

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

This chapter describes the purification and analysis of immune complexes (IC), with the aid of rheumatoid factor-coated tubes. ICs have been detected in sera of patients, with a large variety of autoimmune and malignant diseases. A generally applicable technique for the isolation and analysis of ICs, together with a technique applicable to the identification of glycoprotein components of ICs is described in the chapter. Both are based on the reactivity of ICs with rheumatoid factor (RF). In the first technique, sera, containing ICs, are fractionated on sephacryl S-300 columns, the large molecular-weight fraction is reacted, with RF-coated tubes, and the bound ICs are eluted, radioiodinated, and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography. In the second technique, nonfractionated sera are reacted directly, with RF-coated tubes, the bound ICs are eluted and analyzed, by SDS-PAGE, and the glycoproteins are identified, by their reactivity, with radioiodinated lectins. It is necessary to change the model system, after it became evident that the tetanus toxoid tended to react nonspecifically with human serum proteins. The molecular aggregates, thus, formed could not be distinguished from toxoid complexed, with antibodies, in the fractions, separated by gel filtration.

Original languageEnglish (US)
Pages (from-to)664-675
Number of pages12
JournalMethods in Enzymology
Volume74
Issue numberC
DOIs
StatePublished - Jan 1 1981
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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